Treatment of obesity with G protein-coupled receptor 75 (GPR75) inhibitors

Abstract

The present disclosure provides methods for treating a subject with obesity, methods for identifying a subject at high risk for developing obesity, methods for detecting human G protein-coupled receptor 75 variant nucleic acid molecules and variant polypeptides, and GPR75 variant nucleic acid molecules and variant polypeptides.

Description

[Technical Field]

[0001] Reference to sequence listings This application includes a sequence listing that was electronically filed on June 20, 2021, as a text file of size 533 kilobytes, under the name 18923804802SEQ. This sequence listing is incorporated herein by reference.

[0002] This disclosure generally relates to the treatment of obese subjects with G protein-coupled receptor 75 (GPR75) inhibitors, methods for identifying subjects at high risk of developing obesity, methods for detecting GPR75 variant nucleic acid molecules and variant polypeptides, and GPR75 variant nucleic acid molecules and GPR75 variant polypeptides. [Background technology]

[0003] Obesity and its cardiovascular and metabolic complications, particularly type 2 diabetes and coronary artery disease, are major causes of morbidity and mortality worldwide. There is a significant unmet medical need for safe and effective weight loss approaches.

[0004] Lifestyle interventions related to diet and physical activity are the first-line treatment for obesity and overweight, but their effectiveness can be limited, and weight often returns to normal. Bariatric surgery can be highly effective in reducing weight in severely obese or high-risk patients, but its use is limited by its invasive nature, cost, and the risk of perioperative adverse events, including perioperative mortality. While some medications have shown weight-reducing effects, drug therapy for obesity is limited by barriers to treatment, including the slight weight loss induced by most medications, the side effect profiles of some drugs, contraindications, low adherence rates, and underprescription.

[0005] GPR75 is a member of the G protein-coupled receptor family. GPRs are cell surface receptors that activate guanine nucleotide-binding proteins upon ligand binding. GPR75 is activated by the chemokine CCL5 / RANTES. GPR75 is likely coupled to the heterotrimeric Gq protein and, upon activation, stimulates inositol triphosphate production and calcium recruitment. GPR75, along with CCL5 / RANTES, may play a role in neuronal survival through the activation of downstream signaling pathways including PI3, Akt, and MAP kinases. CCL5 / RANTES can also regulate insulin secretion by pancreatic islet cells through activation of this receptor. [Overview of the Initiative]

[0006] This disclosure provides a method for treating a subject having obesity, the method comprising administering a G protein-coupled receptor 75 (GPR75) inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0007] This disclosure also provides a method for treating a subject having an overweight body weight, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0008] This disclosure also provides a method for treating subjects with elevated BMI, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0009] This disclosure also provides a method for treating subjects with elevated body fat mass, body fat percentage, or body fat volume, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0010] This disclosure also provides a method for treating a subject who has ingested excessive amounts of food, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0011] This disclosure also provides a method for treating a subject to prevent weight gain or maintain weight loss, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0012] The Disclosure also provides a method for treating a subject with a therapeutic agent for treating or suppressing obesity, wherein the subject is obese, and the method comprises the steps of: determining whether the subject has a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide by obtaining or having obtained a biological sample from the subject and performing or having performed a sequence analysis on the biological sample to determine whether the subject has a genotype containing a GPR75 missense variant nucleic acid molecule; administering or continuing to administer a therapeutic agent for treating or suppressing obesity and / or a GPR75 inhibitor at a standard dose to the subject, which is a GPR75 reference; and administering or continuing to administer the therapeutic agent for treating or suppressing obesity and / or a GPR75 inhibitor at the same or a lower standard dose to the subject, which is heterozygous for the GPR75 missense variant nucleic acid molecule, wherein the presence of a genotype containing a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide indicates that the subject has a reduced risk of developing obesity.

[0013] This disclosure also provides a method for identifying subjects at high risk of developing obesity, the method comprising determining, or having determined, the presence or absence of a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide in a biological sample obtained from the subject, wherein if the subject is a GPR75 reference, the subject is at high risk of developing obesity, and if the subject is heterozygous or homozygous for the GPR75 missense variant nucleic acid molecule, the subject is at low risk of developing obesity.

[0014] This disclosure also provides a method for detecting human GPR75 variant nucleic acid molecules in a subject, wherein the method assays a sample obtained from the subject and determines whether the nucleic acid molecule in the sample lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, contains adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, contains thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, or contains A at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1 A genome nucleic acid molecule containing a nucleotide sequence that lacks AAG tetranucleotide or its complement, includes an insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, or includes guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99, lacks CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, or lacks CCAGUAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 8 It contains, lacks the CCAGUAG heptanucleotide or its complement at positions 361-367 as described in SEQ ID NO: 9, lacks the CCAGUAG heptanucleotide or its complement at positions 600-606 as described in SEQ ID NO: 10, contains adenine or its complement at position 556 as described in SEQ ID NO: 12, contains adenine or its complement at position 457 as described in SEQ ID NO: 17, contains adenine or its complement at position 378 as described in SEQ ID NO: 22, and SEQ ID NO: 27 It contains adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 13, uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, and lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7.Lacking AAAG tetranucleotide or its complement at positions 820-823 as described in SEQ ID NO: 8, Lacking AAAG tetranucleotide or its complement at positions 741-744 as described in SEQ ID NO: 9, Lacking AAAG tetranucleotide or its complement at positions 980-983 as described in SEQ ID NO: 10, Including an insertion of uracil or its complement at position 1,410 as described in SEQ ID NO: 15, Including an insertion of uracil or its complement at position 1,311 as described in SEQ ID NO: 20 This includes insertion of a body, insertion of uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, insertion of uracil or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 30, insertion of guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, insertion of guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, insertion of guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or insertion of a body at the position corresponding to position 891 as described in SEQ ID NO: 103 An mRNA molecule having a nucleotide sequence containing anine or its complement, or a cDNA molecule generated from an mRNA molecule, which lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, and is described in SEQ ID NO: 34. Lacking the CCAGTAG heptanucleotide or its complement at positions 600-606 as described above, containing adenine or its complement at position 556 as described in SEQ ID NO: 36, containing adenine or its complement at position 457 as described in SEQ ID NO: 41, containing adenine or its complement at position 378 as described in SEQ ID NO: 46, containing adenine or its complement at position 617 as described in SEQ ID NO: 51, containing thymine or its complement at position 910 as described in SEQ ID NO: 37Contains thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, contains thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, contains thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 741-744 as described in SEQ ID NO: 33, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 980-983 as described in SEQ ID NO: 34, the position corresponding to position 1,410 as described in SEQ ID NO: 39 The present invention provides a method for determining whether a cDNA molecule has a nucleotide sequence that includes an insertion of thymine or its complement at position 1,311 as described in SEQ ID NO: 44, an insertion of thymine or its complement at position 1,232 as described in SEQ ID NO: 49, an insertion of thymine or its complement at position 1,471 as described in SEQ ID NO: 54, an insertion of guanine or its complement at position 830 as described in SEQ ID NO: 104, an insertion of guanine or its complement at position 731 as described in SEQ ID NO: 105, an insertion of guanine or its complement at position 652 as described in SEQ ID NO: 106, or an insertion of guanine or its complement at position 891 as described in SEQ ID NO: 107.

[0015] The disclosure also provides a method for detecting the presence of human GPR75 Ala110fs, Ala116Thr, Tyr207Cys, Gln234Stop, Arg236fs, or Cys400fs variant polypeptides, comprising performing an assay on a sample obtained from a subject and determining whether the GPR75 protein in the sample contains SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, or SEQ ID NO: 108.

[0016] This disclosure also provides isolated change-specific probes or change-specific primers comprising at least about 15 nucleotides, wherein the change-specific probe or change-specific primer comprises a nucleotide sequence complementary to a portion of the nucleotide sequence encoding the human GPR75 polypeptide, the portion of which corresponds to or is complementary to positions 5,540-5,546 as described in SEQ ID NO: 2, positions 539-545 as described in SEQ ID NO: 11, positions 440-446 as described in SEQ ID NO: 16, and the sequence The positions corresponding to positions 361-367 in sequence number 21 or their complements, the positions corresponding to positions 600-606 in sequence number 26 or their complements, the positions corresponding to positions 539-545 in sequence number 35 or their complements, the positions corresponding to positions 440-446 in sequence number 40 or their complements, the positions corresponding to positions 361-367 in sequence number 45 or their complements, or the positions corresponding to positions 600-606 in sequence number 50 or their complements, the positions corresponding to positions 5,920-5,923 in sequence number 5 or their complements , the position corresponding to positions 919-922 in Sequence ID No. 14 or its complement, the position corresponding to positions 820-823 in Sequence ID No. 19 or its complement, the position corresponding to positions 741-744 in Sequence ID No. 24 or its complement, the position corresponding to positions 980-983 in Sequence ID No. 29 or its complement, the position corresponding to positions 919-922 in Sequence ID No. 38 or its complement, the position corresponding to positions 820-823 in Sequence ID No. 43 or its complement, the position corresponding to positions 741-744 in Sequence ID No. 48 or its complement, and This corresponds to the position or complement of position 980-983 described in SEQ ID NO: 53, or the position or complement of position 6,411 described in SEQ ID NO: 6, the position or complement of position 1,410 described in SEQ ID NO: 15, the position or complement of position 1,311 described in SEQ ID NO: 20, the position or complement of position 1,232 described in SEQ ID NO: 25, the position or complement of position 1,471 described in SEQ ID NO: 30, the position or complement of position 1,410 described in SEQ ID NO: 39, and the position 1,This includes the position corresponding to position 311 or its complement, the position corresponding to position 1,232 as described in SEQ ID NO: 49 or its complement, the position corresponding to position 1,471 as described in SEQ ID NO: 54, the position corresponding to position 5,831 as described in SEQ ID NO: 61, the position corresponding to position 830 as described in SEQ ID NO: 100, the position corresponding to position 731 as described in SEQ ID NO: 101, the position corresponding to position 652 as described in SEQ ID NO: 102, the position corresponding to position 891 as described in SEQ ID NO: 103, the position corresponding to position 830 as described in SEQ ID NO: 104, the position corresponding to position 731 as described in SEQ ID NO: 105, the position corresponding to position 652 as described in SEQ ID NO: 106, or the position corresponding to position 891 as described in SEQ ID NO: 107.

[0017] This disclosure also provides a molecular complex comprising a change-specific primer or change-specific probe hybridized with a genomic nucleic acid molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, wherein the change-specific primer or probe hybridizes with a nucleotide or its complement at the position corresponding to positions 5,539-5,540 as described in SEQ ID NO: 2, adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, a nucleotide or its complement at the position corresponding to positions 5,919-5,920 as described in SEQ ID NO: 5, thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, or guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99.

[0018] This disclosure also provides a molecular complex comprising a change-specific primer or change-specific probe hybridized with an mRNA molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, wherein the change-specific primer or probe is a nucleotide or its complement at the position corresponding to positions 538-539 as described in SEQ ID NO: 11, a nucleotide or its complement at the position corresponding to positions 439-440 as described in SEQ ID NO: 16, a nucleotide or its complement at the position corresponding to positions 360-361 as described in SEQ ID NO: 21, a nucleotide or its complement at the position corresponding to positions 599-600 as described in SEQ ID NO: 26, adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22, adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, and SEQ ID NO: 13 Uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 18, uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 23, uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 28, nucleotide or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, nucleotide or its complement at the position corresponding to positions 918-919 as described in SEQ ID NO: 14, nucleotide or its complement at the position corresponding to positions 819-820 as described in SEQ ID NO: 19, nucleotide or its complement at the position corresponding to positions 740-741 as described in SEQ ID NO: 24, nucleotide or its complement at the position corresponding to positions 979-980 as described in SEQ ID NO: 29, uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15, uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20, uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, 1,It hybridizes with uracil or its complement at position 471, guanine or its complement at position 830 as described in SEQ ID NO: 100, guanine or its complement at position 731 as described in SEQ ID NO: 101, guanine or its complement at position 652 as described in SEQ ID NO: 102, or guanine or its complement at position 891 as described in SEQ ID NO: 103.

[0019] This disclosure also provides a molecular complex comprising a change-specific primer or change-specific probe hybridized with a cDNA molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, wherein the change-specific primer or probe is a nucleotide or its complement at the position corresponding to positions 538-539 as described in SEQ ID NO: 35, a nucleotide or its complement at the position corresponding to positions 439-440 as described in SEQ ID NO: 40, a nucleotide or its complement at the position corresponding to positions 360-361 as described in SEQ ID NO: 45, a nucleotide or its complement at the position corresponding to positions 599-600 as described in SEQ ID NO: 50, adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, sequence Thymine or its complement at the position corresponding to position 910 as described in number 37, thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, nucleotide or its complement at the positions corresponding to positions 918-919 as described in SEQ ID NO: 38, nucleotide or its complement at the positions corresponding to positions 819-820 as described in SEQ ID NO: 43, nucleotide or its complement at the positions corresponding to positions 740-741 as described in SEQ ID NO: 48, nucleotide or its complement at the positions corresponding to positions 979-980 as described in SEQ ID NO: 53, thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, 1,It hybridizes with thymine or its complement at position 471, guanine or its complement at position 830 as described in SEQ ID NO: 104, guanine or its complement at position 731 as described in SEQ ID NO: 105, guanine or its complement at position 652 as described in SEQ ID NO: 106, or guanine or its complement at position 891 as described in SEQ ID NO: 107.

[0020] The disclosure also provides an isolated nucleic acid molecule or its complement comprising a nucleotide sequence encoding a human GPR75 polypeptide, the polypeptide comprising a frameshift starting at the position corresponding to position 110 as described in SEQ ID NO: 56, a frameshift starting at the position corresponding to position 236 as described in SEQ ID NO: 59, or a frameshift starting at the position corresponding to position 400 as described in SEQ ID NO: 60.

[0021] The disclosure also provides an isolated genomic nucleic acid molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, the nucleotide sequence containing either a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 5,540–5,546 as described in SEQ ID NO: 1, or an AAAG tetranucleotide or its complement at the position corresponding to positions 5,920–5,923 as described in SEQ ID NO: 1, or an insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6.

[0022] This disclosure also provides an isolated mRNA molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, wherein this nucleotide sequence lacks a CCAGUAG heptanucleotide or its complement at positions 539-545 as described in SEQ ID NO: 7, or at positions 440-446 as described in SEQ ID NO: 8, or at positions 361-367 as described in SEQ ID NO: 9, or at positions 600-606 as described in SEQ ID NO: 10, or at positions 919-922 as described in SEQ ID NO: 7. It may be: lacking AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 8; lacking AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 9; lacking AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 10; including an insertion of uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15; including an insertion of uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20; including an insertion of uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25; or including an insertion of uracil or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 30.

[0023] This disclosure also provides an isolated cDNA molecule containing a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence lacks a CCAGTAG heptanucleotide or its complement at positions 539-545 as described in SEQ ID NO: 31, or at positions 440-446 as described in SEQ ID NO: 32, or at positions 361-367 as described in SEQ ID NO: 33, or at positions 600-606 as described in SEQ ID NO: 34, or lacks an AAAG tetranucleotide or its complement at positions 919-922 as described in SEQ ID NO: 31. It contains, or lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, or lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, or includes an insertion of thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, or includes an insertion of thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, or includes an insertion of thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or includes an insertion of thymine or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 54.

[0024] The disclosure also provides an isolated human GPR75 polypeptide having an amino acid sequence at least about 90% identical to SEQ ID NO: 56, lacking the amino acids at positions 110-540 described in SEQ ID NO: 55; an isolated human GPR75 polypeptide having an amino acid sequence at least about 90% identical to SEQ ID NO: 59, lacking the amino acids at positions 236-540 described in SEQ ID NO: 55; or an isolated human GPR75 polypeptide having an amino acid sequence at least about 90% identical to SEQ ID NO: 60, lacking the amino acids at positions 400-540 described in SEQ ID NO: 55.

[0025] This disclosure also provides therapeutic agents for treating or suppressing obesity for use in the treatment of the subject, which is a genomic nucleic acid molecule having a nucleotide sequence encoding the human GPR75 polypeptide, wherein the nucleotide sequence lacks the CCAGUAG heptanucleotide or its complement at positions 5,540-5,546 as described in SEQ ID NO: 1, includes adenine or its complement at position 5,557 as described in SEQ ID NO: 3, includes thymine or its complement at position 5,911 as described in SEQ ID NO: 4, lacks the AAAG tetranucleotide or its complement at positions 5,920-5,923 as described in SEQ ID NO: 1, includes an insertion of thymine or its complement at position 6,411 as described in SEQ ID NO: 6, or includes guanine or its complement at position 5,831 as described in SEQ ID NO: 99, and is a genomic nucleic acid molecule having a nucleotide sequence encoding the human GPR75 polypeptide, wherein the nucleotide sequence lacks the CCAGUAG heptanucleotide or its complement at positions 539-545 as described in SEQ ID NO: 7 CCAGUAG heptanucleotides at positions 440-446 in SEQ ID NO: 8 that lack their complement, or CCAGUAG heptanucleotides at positions 361-367 in SEQ ID NO: 9 that lack their complement, or CCAGUAG heptanucleotides at positions 600-606 in SEQ ID NO: 10 that lack their complement, or adenine at position 556 in SEQ ID NO: 12, or adenine at position 457 in SEQ ID NO: 17 that contains its complement. Or including its complement, including adenine at the position corresponding to position 378 as described in SEQ ID NO: 22 or its complement, including adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, including uracil at the position corresponding to position 910 as described in SEQ ID NO: 13, including uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, including uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, including uracil at the position corresponding to position 971 as described in SEQ ID NO: 28,Lacking AAAG tetranucleotide or its complement at positions 919-922 as described in SEQ ID NO: 7, Lacking AAAG tetranucleotide or its complement at positions 820-823 as described in SEQ ID NO: 8, Lacking AAAG tetranucleotide or its complement at positions 741-744 as described in SEQ ID NO: 9, Lacking AAAG tetranucleotide or its complement at positions 980-983 as described in SEQ ID NO: 10, Lacking U at position 1,410 as described in SEQ ID NO: 15 Includes insertion of uracil or its complement, including insertion of uracil or its complement at position 1,311 as described in SEQ ID NO: 20, includes insertion of uracil or its complement at position 1,232 as described in SEQ ID NO: 25, includes insertion of uracil or its complement at position 1,471 as described in SEQ ID NO: 30, includes insertion of guanine or its complement at position 830 as described in SEQ ID NO: 100, includes insertion of guanine or its complement at position 731 as described in SEQ ID NO: 101, and SEQ ID NO: 102 An mRNA molecule containing guanine or its complement at the position corresponding to position 652, or containing guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103, or a cDNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, or the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32. Lacking complement, lacking the CCAGTAG heptanucleotide or its complement at positions 361-367 as described in SEQ ID NO: 33, lacking the CCAGTAG heptanucleotide or its complement at positions 600-606 as described in SEQ ID NO: 34, containing adenine or its complement at position 556 as described in SEQ ID NO: 36, containing adenine or its complement at position 457 as described in SEQ ID NO: 41, containing adenine or its complement at position 378 as described in SEQ ID NO: 46,Contains adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, contains thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, contains thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, contains thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, contains thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 741-744 as described in SEQ ID NO: 33, and also contains AAAG tetranucleotide at the positions corresponding to positions 980-983 as described in SEQ ID NO: 34. The present invention provides a therapeutic agent for treating or suppressing obesity in subjects having a cDNA molecule that lacks its complement, includes an insertion of thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, an insertion of thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, an insertion of thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, an insertion of thymine or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 54, an insertion of guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, an insertion of guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, an insertion of guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or an insertion of guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107.

[0026] This disclosure also relates to a genomic nucleic acid molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence lacks a CCAGTAG heptanucleotide or its complement at positions 5,540-5,546 as described in SEQ ID NO: 1, contains adenine or its complement at position 5,557 as described in SEQ ID NO: 3, contains thymine or its complement at position 5,911 as described in SEQ ID NO: 4, and contains an AAAG tetranucleotide or its complement at positions 5,920-5,923 as described in SEQ ID NO: 1. A genome nucleic acid molecule, an mRNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, which includes an insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, or includes guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99, wherein the nucleotide sequence lacks the CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, or the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8 Lack of ocidole or its complement; lacking CCAGUAG heptanucleotide or its complement at positions 361-367 as described in SEQ ID NO: 9; lacking CCAGUAG heptanucleotide or its complement at positions 600-606 as described in SEQ ID NO: 10; containing adenine or its complement at position 556 as described in SEQ ID NO: 12; containing adenine or its complement at position 457 as described in SEQ ID NO: 17; containing adenine or its complement at position 378 as described in SEQ ID NO: 22 , containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, containing uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, containing uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, containing uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, containing uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, lacking AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7.The following are examples of sequences of sequences: 10, 15, and 20, all of which include: 1. Lacking AAAG tetranucleotide or its complement at positions 820-823 as described in Sequence ID No. 8; 2. Lacking AAAG tetranucleotide or its complement at positions 741-744 as described in Sequence ID No. 9; 3. Lacking AAAG tetranucleotide or its complement at positions 980-983 as described in Sequence ID No. 10; 4. Insertion of uracil or its complement at position 1,311 as described in Sequence ID No. 20. This includes, including, the insertion of uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, the insertion of uracil or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 30, the insertion of guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, the insertion of guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, the insertion of guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or the insertion of guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103 An mRNA molecule containing a complement of, or a cDNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, SEQ ID NO: Lacking the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in 34, containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, containing adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, containing adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, containing thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37Contains thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, contains thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, contains thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 741-744 as described in SEQ ID NO: 33, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 980-983 as described in SEQ ID NO: 34, corresponding to position 1,410 as described in SEQ ID NO: 39 The present invention provides a GPR75 inhibitor for use in treating obesity in subjects having a cDNA molecule, including an insertion of thymine or its complement at a position corresponding to position 1,311 as described in SEQ ID NO: 44, an insertion of thymine or its complement at a position corresponding to position 1,232 as described in SEQ ID NO: 49, an insertion of thymine or its complement at a position corresponding to position 1,471 as described in SEQ ID NO: 54, an insertion of guanine or its complement at a position corresponding to position 830 as described in SEQ ID NO: 104, an insertion of guanine or its complement at a position corresponding to position 731 as described in SEQ ID NO: 105, an insertion of guanine or its complement at a position corresponding to position 652 as described in SEQ ID NO: 106, or an insertion of guanine or its complement at a position corresponding to position 891 as described in SEQ ID NO: 107.

[0027] The accompanying drawings, which are incorporated herein and constitute part of herein, illustrate some of the features of this disclosure. This patent or application document includes at least one drawing drawn in color. Copies of this patent or patent application publication, including the color drawing(s), will be provided by the Patent Office upon request and payment of the necessary fees. [Brief explanation of the drawing]

[0028] [Figure 1]This shows the baseline characteristics of individuals included in exome-wide association studies. Abbreviations: UKB: UK Biobank, GHS: Geisinger Health System, MCPS: Mexico City Prospective Study, SD: Standard deviation, N: Number of participants, WHO: World Health Organization, IQR: Interquartile range, kg / m2: Kilograms per square meter, mg / dL: Milligrams per deciliter, mmHg: Millimeters of mercury. [Figure 2] The results of the association of GPR75 with body mass index in the UKB, GHS, and MCPS cohorts are presented. Abbreviations: CI: confidence interval, SD: standard deviation, BMI: body mass index, AAF: alternative allele frequency, RR: reference-reference genotype, RA: reference-alternative heterozygous genotype, AA: alternative-alternative homozygous genotype, pLOF: predicted loss of function, UKB: UK Biobank, GHS: Geisinger Health System MyCode study, MCPS: Mexico City Prospective Study. [Figure 3]This table shows the association between GPR75 and body mass index in exome-wide gene loading analysis. The table lists the association statistics for the GPR75 gene where gene loading of rare pLOF variants was associated with body mass index at the exome-wide level with statistical significance (p<3.6 × 10⁻⁷). The analysis was performed on 645,626 participants from the UKB, GHS, and MCPS studies. Genomic coordinates represent chromosomal and physical locations in base pairs, in accordance with the International Human Genome Reference Sequence (Genome Reference Consortium Human Build) 38. Abbreviations: CI: Confidence interval, SD: Standard deviation, BMI: Body mass index, AAF: Alternate allele frequency, RR: Reference-reference genotype, RA: Reference-alternative heterozygous genotype, AA: Alternate-alternative homozygous genotype, pLOF: Predicted loss of function, Missense (1 / 5): Missense variant predicted to be harmful by at least one of five in silico prediction algorithms, Missense (5 / 5): Missense variant predicted to be harmful by five of five in silico prediction algorithms. [Figure 4A] This panel shows protein truncation variants in GPR75 associated with decreased human body mass index. Panel A shows a linear model of the GPR75 protein and its domains (top, intracellular and extracellular domains (yellow), transmembrane domain (orange)), the distribution of 46 predicted loss-of-function variants found by exome sequencing on the GPR75 protein (center), and the distribution of BMI (standardized units) among heterozygous carriers of each variant (bottom). In the bottom subpanel, the blue horizontal line shows the mean BMI of non-carriers, and the red horizontal line shows the overall mean BMI adjusted for covariates in carriers of any predicted loss-of-function gene variant of GPR75. [Figure 4B]This panel shows protein truncation variants in GPR75 associated with decreased human body mass index. Panel B shows a meta-analysis of the association between predicted loss-of-function variants in GPR75 and BMI in the exploratory and supplemental cohorts. Abbreviations: CI: confidence interval, RR: reference-reference genotype, RA: reference-alternative heterozygous genotype, AA: alternative-alternative homozygous genotype, DHS: Dallas Heart Study, SINAI: Mount Sinai BioMe cohort, DUKE: Duke Catheterization Genetics cohort, TAICHI: Taiwanese Chinese from the Taiwan Metabochip Consortium, PMBB: University of Pennsylvania Medicine BioBank, MALMO: Malmo Diet and Cancer Study, AFR: African, AMR: American, EAS: East Asian, EUR: European, SAS: South Asian. MCPS included mixed-race American individuals. [Figure 5] This report shows the association between GPR75 pLOF variants and body mass index within age and sex subgroups. Abbreviations: Confidence interval: CI, Standard deviation: SD, Body mass index: BMI, Alternate allele frequency: AAF, P-value: p, Reference-reference genotype: RR, Reference-alternative genotype: RA, Alternate-alternative genotype: AA, Kilograms per square meter: kg / m2, Predicted loss of function: pLOF. [Figure 6] This study shows the association of the GPR75 pLOF variant with obesity risk. Abbreviations: OR: odds ratio, CI: confidence interval, P-value: p, AAF: alternative allele frequency, reference-reference genotype: RR, reference-alternative genotype: RA, alternative-alternative genotype: AA, pLOF: predicted loss of function. Results are from a meta-analysis of UKB, GHS, and MCPS studies. [Figure 7]This shows the distribution of body mass index categories for individuals who are carriers and non-carriers of predicted loss-of-function variants in GPR75 or MC4R. It also shows the distribution of heterozygous carriers (top), non-carriers (center), and heterozygous carriers (bottom) of predicted loss-of-function variants in GPR75 and MC4R, according to World Health Organization classification, for the UKB, GHS, and MCPS cohorts. [Figure 8] This study shows the association between self-reported being underweight compared to the average body size at age 10 in the UKB and the GPR75 pLOF variant. Abbreviations: Predicted loss of function: pLOF, UKB: UKB, Surrogate allele frequency: AAF, Confidence interval: CI, Odds ratio: OR, P-value: p, Reference-reference genotype: RR, Reference-surrogate genotype: RA, Surrogate-surrogate genotype: AA. [Figure 9-1] This study shows the association between indices of body fat mass and lean body mass estimated by bioelectrical impedance analysis and pLOF variants in GPR75 and MC4R. Association analyses were performed on 423,418 participants in the UK Biobank study who underwent whole-exome sequencing and bioelectrical impedance measurements. Abbreviations: pLOF: predicted loss of function, kg: kilograms, CI: confidence interval. [Figure 9-2] Same as above. [Figure 10] This study shows the association between cardiovascular and metabolic phenotypes and pLOF gene variants in GPR75 in the UKB, GHS, and MCPS studies. Abbreviations: P-value: p, SD: standard deviation, CI: confidence interval, pLOF: predicted loss of function, AAF: surrogate allele frequency, RR: reference-reference genotype, RA: reference-surrogate heterozygous genotype, AA: surrogate-surrogate homozygous genotype, hemoglobin A1C: HbA1c, aspartate transaminase: AST, alanine transaminase: ALT, high-density lipoprotein cholesterol: HDL-C, low-density lipoprotein cholesterol: LDL-C, milligrams per deciliter: mg / dL, millimeters of mercury: mmHg, units per liter: U / L. [Figure 11] The GPR75, ASB3, and GPR75-ASB3 genes are shown. The figure shows the gene models and chromosomal locations of the GPR75, GPR75-ASB3, and ASB3 genes. GPR75 shares exon 1, which contains the non-coding sequence, with the GPR75-ASB3 readthrough gene. Exon 2 of GPR75, which contains the entire coding sequence, is exclusive to the GPR75 gene and is not shared with any other gene. ASB3 and GPR75-ASB3 share some exons with each other but not with GPR75. The underlying description is from an Ensembl region plot (version 85). [Figure 12-1] This table shows the predicted loss-of-function (pLOF) variants of GPR75 identified by exome sequencing. The table lists the predicted loss-of-function (pLOF) variants of the GPR75 gene found by exome sequencing that contributed to gene loading analysis. An INFO score of less than 0.3 is generally considered extremely low quality. Abbreviations: chromosome, location, reference, alternative: CPRA, alternative allele frequency: AAF, complementary DNA: cDNA, Human Genome Variation Society: HGVS. [Figure 12-2] Same as above. [Figure 12-3] Same as above. [Figure 13-1] This study indicates that the loading of rare non-synonymous variants in ASB3 or GPR75-ASB3 is not associated with BMI. Abbreviations: Alternate allele frequency: AAF, Confidence interval: CI, Standard deviation: SD, Body mass index: BMI, Kilograms per square meter: kg / m2, P-value: p, Reference-reference genotype: RR, Reference-alternative genotype: RA, Alternate-alternative genotype: AA, Predicted loss of function: pLOF. [Figure 13-2] Same as above. [Figure 14]This shows the association between pLOF variant loadings in GPR75 and BMI, after adjusting for ASB3, GPR75-ASB3, and common variant genotypes within the region. Abbreviations: Confidence interval: CI, Standard deviation: SD, Body mass index: BMI, Kilograms per square meter: kg / m2, P-value: p, Alternate allele frequency: AAF, Reference-reference genotype: RR, Reference-alternative genotype: RA, Alternate-alternative genotype: AA, Predicted loss of function: pLOF. *Standardized covariates included all fine-mapped common variants from GWAS analysis, including rs59428052 near ASB3. [Figure 15-1] This table shows common variants associated with BMI at the GPR75 locus. The table lists 26 common variants associated with BMI at a genome-wide statistical significance threshold (p<5×10⁻⁸) within the bilateral 500kb of GPR75 in Europeans. Variants are annotated to their nearest neighbor genes and are also annotated as being eQTL sentinels or non-synonymous coding variants with LD (R^2>0.8). In mixed-race Americans, no common variants associated with BMI at genome-wide significance were found at the GPR75 locus. Abbreviations: chromosome, location, reference, alternative: CPRA, alternative allele frequency: AAF, posterior probability of causality: PPA, linkage disequilibrium: LD, quantitative expression trait locus: eQTL. [Figure 15-2] Same as above. [Figure 15-3] Same as above. [Figure 16-1] This section shows the association between common variants at the GPR75 locus and BMI. The left panel shows the results of GWAS analyses of imputed common variants in European individuals of UKB and GHS, while the right panel shows the results of GWAS analyses of the same variants in mixed-race Americans from the MCPS cohort. The sentinel variant (rs59428052) from the European GWAS is highlighted in the left panel. No significant genome-wide association was found in mixed-race Americans (p<5×10⁻⁸). [Figure 16-2] Same as above. [Figure 17-1]This table shows the association between body mass index and pLOF gene variants in GPR75 in sensitivity analyses. The table includes leave-one-out analyses, which exclude one gene variant at a time, and analyses (bottom row) that exclude variants associated with BMI reduction in individual variant analyses. Abbreviations: CI: confidence interval, SD: standard deviation, BMI: body mass index, p: p-value, pLOF: predicted loss of function. [Figure 17-2] Same as above. [Figure 17-3] Same as above. [Figure 17-4] Same as above. [Figure 18] This table shows the predicted loss-of-function variants in GPR75 associated with BMI in individual variant analyses. It also includes association statistics for GPR75 pLOF variants that were included in gene loading analyses and were individually associated with BMI in inverse variance-weighted meta-analysis (p<0.05). Abbreviations: AAF: Alternate allele frequency, CI: Confidence interval, SD: Standard deviation, BMI: Body Mass Index, p: P-value, Frameshift: fs, pLOF: Predicted loss of function. [Figure 19A] This panel shows in vitro expression studies of two predicted loss-of-function gene variants in GPR75. Panel A shows the results of quantitative reverse transcription polymerase chain reaction experiments measuring GPR75 mRNA levels. GPR75 expression levels were calculated relative to the beta-actin gene. The values ​​represent the mean and standard deviation of three technical replicates, each representing one of three biological replicates performed for each condition. [Figure 19B] This panel shows in vitro expression studies of two predicted loss-of-function gene variants in GPR75. Panel B shows Western blotting analysis at the GPR75 protein level. The GPR75 Ala110fs and Gln234* protein products correspond to predicted molecular weights of 14 kDa and 25 kDa, respectively. The results represent three biological replicates. [Figure 19C]This panel shows in vitro expression studies of two predicted loss-of-function gene variants in GPR75. Panel C shows immunofluorescence staining experiments describing the cellular localization of GPR75. The upper image shows intracellular staining performed by membrane permeabilization, and the lower image shows plasma membrane localization (impermeabilized cell membrane). [Figure 19D] This panel shows in vitro expression studies of two predicted loss-of-function gene variants in GPR75. Panel D shows flow cytometry analysis of GPR75 cell surface expression. Identified cell populations are expressed as percentages (%) of HA-TAG GPR75-positive live cells. Values ​​represent the mean and standard deviation of four biological replicates for each condition. All experiments were performed on HEK293 cells transfected with green fluorescent protein control plasmid (control), GPR75-wild type (GPR75), GPR75-Ala110fs, or GPR75-Gln234* plasmid. Abbreviations: SSC: Lateral scattering, HA: Hemagglutinin tag. [Figure 20] This shows the association between N-terminal truncation gene variants and C-terminal truncation gene variants of GPR75 and BMI. Abbreviations: CI: confidence interval, SD: standard deviation, BMI: body mass index, p: p-value, AAF: alternative allele frequency, RR: reference-reference genotype, RA: reference-alternative heterozygous genotype, AA: alternative-alternative homozygous genotype, kg / m2: kilograms per square meter. [Figure 21] This graph shows the average body weight of male (upper panel) and female (lower panel) GPR75 WT (indicated as HIST and WT) and homozygous knockout (indicated as KO) mice. [Figure 22] This shows the percentage difference in body composition between male homozygous knockout (indicated as KO) and wild-type individuals. BMC: Bone mineral content, BMD: Bone mineral density, P: p-value. [Figure 23-1] The mean fat volume and fat volume percentage of GPR75 WT (HIST and WT) male mice and homozygous knockout (indicated by KO) male mice are shown. [Figure 23-2] Same as above. [Figure 23-3] Same as above. [Figure 24A] This study shows body weight gain and metabolic phenotypes in mice with Gpr75 gene deletion during high-fat diet exposure. Panel A shows weekly body weight gain during 14 weeks of high-fat diet exposure. The panel shows results for Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice. The number of mice in each group and analysis is shown in parentheses. Results are expressed as mean ± standard error. Abbreviations: ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are based on two-way ANOVA and Tukey's multiple comparison test. [Figure 24B] This panel shows body weight gain and metabolic phenotype in mice with Gpr75 gene deletion during high-fat diet exposure. Panel B shows changes in fasting blood glucose before and after high-fat diet exposure. The panel shows results for Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice. The number in parentheses indicates the number of mice included in each group and analysis. Results are expressed as mean ± standard error. Abbreviations: ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are based on two-way ANOVA and Tukey's multiple comparison test. [Figure 24C] This panel shows body weight gain and metabolic phenotype during high-fat diet in mice with Gpr75 gene deletion. Panel C shows the results of the glucose tolerance test at the end of 14 weeks of high-fat diet exposure. The panel shows results for Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice. The number in parentheses indicates the number of mice included in each group and analysis. Results are expressed as mean ± standard error. Abbreviations: ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are from two-way ANOVA and Tukey's multiple comparison test. [Figure 24D]This panel shows body weight gain and metabolic phenotypes in mice with Gpr75 gene deletion during high-fat diet exposure. Panel D shows plasma insulin at the end of 14 weeks of high-fat diet exposure. The panel shows results for Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice. The number of mice in each group and analysis is shown in parentheses. Results are expressed as mean ± standard error. Abbreviations: ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are from two-way ANOVA and Tukey's multiple comparison test. [Figure 25A] This panel shows plasma leptin, adiponectin, and leptin-adiponectin ratios in mouse experiments. Panel A shows plasma leptin levels in Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice after exposure to a high-fat diet, expressed as a multiplier difference compared to wild-type mice (set as 1). The absolute level (mean ± standard deviation) for wild-type mice was 208 ± 42 pg / mL. The numbers in parentheses on the X-axis indicate the number of mice included in each group and analysis. Results are expressed as mean ± standard error. ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are from two-way ANOVA and Tukey's multiple comparison test. [Figure 25B] This panel shows plasma leptin, adiponectin, and leptin-adiponectin ratios in mouse experiments. Panel B shows plasma adiponectin levels in Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice after exposure to a high-fat diet, expressed as a multiplier difference compared to wild-type mice (set as 1). The absolute level (mean ± standard deviation) for wild-type mice was 3,911 ± 1,656 ng / mL. The number in parentheses on the X-axis indicates the number of mice included in each group and analysis. Results are expressed as mean ± standard error. ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are from two-way ANOVA and Tukey's multiple comparison test. [Figure 25C]This panel shows plasma leptin, adiponectin, and leptin-adiponectin ratios in mouse experiments. Panel C shows leptin-to-adiponectin ratios, expressed as a ratio, for Gpr75+ / + (WT), Gpr75+ / - (HET), and Gpr75- / - (KO) mice. The number in parentheses on the X-axis indicates the number of mice included in each group and analysis. Results are expressed as mean ± standard error. ns: Not statistically significant. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 are based on two-way ANOVA and Tukey's multiple comparison test. [Figure 26A] This shows body weight, fat mass, lean body mass, bone mass, and blood glucose levels measured in a high-fat diet mouse experiment. The body weight (Panel A) of Gpr75+ / +, Gpr75+ / -, and Gpr75- / - mice (black circles, red squares, and blue triangles, respectively) maintained on solid diet (week 0 or -1) and high-fat diet (HFD) for 9 weeks. *: P<0.05 for Gpr75- / - mice compared to Gpr75+ / + mice. #: P<0.05 for Gpr75+ / - mice compared to Gpr75+ / + mice. $: For each genotype, the data point from the last high-fat diet is P<0.05 compared to the first data point obtained on solid diet before high-fat diet feeding. [Figure 26B-1] This shows body weight, fat mass, lean body mass, bone mass, and blood glucose levels measured in a high-fat diet mouse experiment. Fat mass (top panel B), lean body mass (center panel B), and bone mass (bottom panel B) are shown for Gpr75+ / +, Gpr75+ / -, and Gpr75- / - mice (black circles, red squares, and blue triangles, respectively) maintained on solid diet (week 0 or -1) and high-fat diet (HFD) for 9 weeks. *: P<0.05 for Gpr75- / - mice compared to Gpr75+ / + mice. #: P<0.05 for Gpr75+ / - mice compared to Gpr75+ / + mice. $: For each genotype, the data point from the last high-fat diet is P<0.05 compared to the first data point obtained on solid diet before high-fat diet feeding. [Figure 26B-2] Same as above. [Figure 26B-3] Same as above. [Figure 26C]This shows body weight, fat mass, lean body mass, bone mass, and blood glucose levels measured in a high-fat diet mouse experiment. Blood glucose levels (Panel C) of Gpr75+ / +, Gpr75+ / -, and Gpr75- / - mice (black circles, red squares, and blue triangles, respectively) maintained on solid diet (week 0 or -1) and high-fat diet (HFD) for 9 weeks. *: P<0.05 for Gpr75- / - mice compared to Gpr75+ / + mice. #: P<0.05 for Gpr75+ / - mice compared to Gpr75+ / + mice. $: For each genotype, the data point from the last high-fat diet is P<0.05 compared to the first data point obtained on solid diet before high-fat diet feeding. [Figure 27] This study demonstrates that female GPR75 knockout mice fed solid feed show improved glucose tolerance rather than insulin resistance. [Modes for carrying out the invention]

[0029] Various terms relating to aspects of this disclosure are used throughout this specification and the claims. Unless otherwise specified, such terms shall be given their common meanings in the art. Other terms specifically defined shall be construed in accordance with the definitions set forth herein.

[0030] Unless otherwise expressly provided, none of the methods or embodiments described herein are intended to be construed as requiring their steps to be performed in a specific order. Therefore, unless the claims or description of a method specifically require that the steps be limited to a particular order, no order is intended to be inferred in any way. This also applies to any possible implicit criteria of interpretation, including logical matters concerning the arrangement of steps or workflows, general meanings arising from grammatical construction or punctuation, or the number or types of embodiments described herein.

[0031] As used herein, unless otherwise clearly indicated by the context, the singular forms "a," "an," and "the" refer to multiple objects. Where used herein, the term “approximately” means that the stated figures are approximate and small variations will not significantly affect the practice of the disclosed embodiments. Where figures are used, unless otherwise indicated by the context, the term “approximately” means that the figures may vary by ±10% and remain within the scope of the disclosed embodiments.

[0032] As used herein, the term “including” may, in certain embodiments, be replaced as desired with “consisting of” or “essentially consisting of.” As used herein, the term “isolated” with respect to a nucleic acid molecule or polypeptide means that the nucleic acid molecule or polypeptide is in a state other than its original environment, for example, away from blood and / or animal tissue. In some embodiments, the isolated nucleic acid molecule or polypeptide is substantially free from other nucleic acid molecules or other polypeptides, particularly other nucleic acid molecules or polypeptides of animal origin. In some embodiments, the nucleic acid molecule or polypeptide may be in a highly purified form, i.e., with a purity greater than 95% or greater than 99%. As used in this context, the term “isolated” does not exclude the presence of the same nucleic acid molecule or polypeptide in dimers or alternative physical forms such as phosphorylated or derivatized forms.

[0033] As used herein, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid sequence,” “polynucleotide,” or “oligonucleotide” may include polymeric forms of nucleotides of any length, may include DNA and / or RNA, and may be single-stranded, double-stranded, or multi-stranded. One strand of a nucleic acid also refers to its complement.

[0034] As used herein, the term “subject” includes any animal, including mammals. Mammals include, but are not limited to, livestock (e.g., horses, cattle, pigs), companion animals (e.g., dogs, cats), laboratory animals (e.g., mice, rats, rabbits), and non-human primates (e.g., apes and monkeys). In some embodiments, the subject is a human. In some embodiments, the subject is a patient under the care of a physician.

[0035] Variants in the GPR75 gene associated with a reduced risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake have been identified in accordance with this disclosure. For example, genetic alterations that delete the CCAGTAG heptanucleotide at positions 5,540–5,546 of human GPR75 (see SEQ ID NO: 1), or that change the guanine nucleotide at position 5,557 of human GPR75 (see SEQ ID NO: 1) to adenine, or the tyrosine nucleotide at position 5,911 of human GPR75 (see SEQ ID NO: 1) to thymine, or that delete the AAAG tetranucleotide at positions 5,920–5,923 of human GPR75 (see SEQ ID NO: 1), or that insert a thymine nucleotide at position 6,411 of human GPR75 (see SEQ ID NO: 1), have been shown to reduce the risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake in individuals with such alterations. It is believed that no GPR75 gene or protein variants have any known association with obesity, weight, BMI, body fat mass, body fat percentage, or body fat volume, and / or lean body mass, lean body mass percentage, or lean body mass volume. Overall, the genetic analyses described herein have surprisingly shown that the GPR75 gene, and in particular variants of the GPR75 gene, are associated with a reduced risk of developing obesity and with a decrease in weight, a decrease in BMI, a decrease in body fat mass, body fat percentage, or body fat volume, and / or a decrease in lean body mass, lean body mass percentage, or lean body mass volume. Therefore, subjects who are GPR75 references and at high risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake can be treated to prevent obesity, reduce its symptoms, and / or suppress the onset of symptoms.Accordingly, this disclosure provides a method for leveraging the identification of such variants of a subject to identify or stratify the risk of such subjects developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake, or to diagnose subjects as being at high risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake, thereby enabling the treatment of subjects at risk or subjects with active disease. Furthermore, this disclosure provides isolated GPR75 variant genomic nucleic acid molecules, variant mRNA molecules, and variant cDNA molecules. The specification also provides loss-of-function variant nucleic acid molecules of GPR75 that have been found to be associated with a reduced risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake.

[0036] Further missense variants of the GPR75 gene that may be associated with a reduced risk of developing obesity, overweight, elevated BMI, increased body fat mass, body fat percentage, or body fat volume, and / or excessive food intake include: a substitution of thymine with cytosine at position 53,854,656 on chromosome 2 (according to the GRCh38 / hg38 (December 2013) Human Genome Assembly), resulting in a substitution of glutamine with arginine at the position corresponding to position 34 described in SEQ ID NO: 55, a substitution of guanine with alanine at position 53,854,330 on chromosome 2, resulting in a substitution of arginine with tryptophan at the position corresponding to position 143 described in SEQ ID NO: 55, and a substitution of thymine with cytosine at position 53,854,321 on chromosome 2, resulting in a substitution of methionine with valine at the position corresponding to position 146 described in SEQ ID NO: 55. Substitutions include, but are not limited to, the substitution of cytosine at position 53,854,191 of chromosome 2 with guanine, resulting in the substitution of cysteine ​​with serine at the position corresponding to position 189 as described in SEQ ID NO: 55; the substitution of isoleucine at position corresponding to position 326 as described in SEQ ID NO: 55, resulting in the substitution of alanine at position 53,853,780 of chromosome 2 with guanine; the substitution of isoleucine at position corresponding to position 375 as described in SEQ ID NO: 55, resulting in the substitution of thymine at position 53,853,634 of chromosome 2 with adenine; and the substitution of glutamic acid at position corresponding to position 526 as described in SEQ ID NO: 55, resulting in the substitution of cytosine at position 53,853,181 of chromosome 2 with lysine (and thus may be included in any of the embodiments described herein).

[0037] Further variants of the GPR75 gene may be associated with an increased risk of developing obesity, overweight, elevated BMI, increased body fat mass, body fat percentage, or body fat volume, and / or excessive food intake in subjects, and these may be considered gain-of-function (GOF) variants. Such GOFs may include a substitution of thymine for adenine at position 53,854,437 on chromosome 2, resulting in a substitution of tyrosine for phenylalanine at the position corresponding to position 107 as described in SEQ ID NO: 55, and a substitution of phenylalanine for guanine at position 53,853,697 on chromosome 2, resulting in a substitution of phenylalanine for leucine at the position corresponding to position 354 as described in SEQ ID NO: 55 (in accordance with the GRCh38 / hg38 (December 2013) Human Genome Assembly). Therefore, subjects with any of these variants can be treated with GPR75 inhibitors. Such individuals can be assessed for the risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake by detecting the presence of GOF variants.

[0038] For the purposes of this disclosure, any particular human being can be classified as having one of three GPR75 genotypes: i) GPR75 reference, ii) heterozygous for a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide, or iii) homozygous for a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide. If a human being does not have a copy of the GPR75 missense variant nucleic acid molecule, that human being is GPR75 reference. If a human being has a single copy of the GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide, that human being is heterozygous for a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide. As used herein, a GPR75 missense variant nucleic acid molecule is any GPR75 nucleic acid molecule (e.g., a genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) encoding a GPR75 polypeptide having partial loss of function, complete loss of function, predicted partial loss of function, or predicted complete loss of function. A human having a GPR75 polypeptide with partial loss of function (or predicted partial loss of function) is low phenotypic for GPR75. A predicted loss of function variant nucleic acid molecule of GPR75 may be any nucleic acid molecule encoding GPR75 Ala110fs, Ala116Thr, Tyr207Cys, Gln234Stop, Arg236fs, or Cys400fs. A GPR75 missense variant nucleic acid molecule may also be Lys404 * This could be any nucleic acid molecule encoding Ser219fs, or it could be c.-110+1G>A. If a human has two copies of the GPR75 missense variant nucleic acid molecule, that human is homozygous for the GPR75 missense variant nucleic acid molecule encoding the predicted loss-of-function GPR75 polypeptide.

[0039] For subjects genotyped or determined to be GPR75 reference, such subjects are at high risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake. For subjects genotyped or determined to be heterozygous for either a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide, such subjects can be treated with a GPR75 inhibitor.

[0040] In any of the embodiments described herein, the GPR75 missense variant nucleic acid molecule may be any GPR75 nucleic acid molecule (e.g., a genomic nucleic acid molecule, an mRNA molecule, or a cDNA molecule) encoding a GPR75 polypeptide having partial loss of function, complete loss of function, predicted partial loss of function, or predicted complete loss of function. For example, the GPR75 missense variant nucleic acid molecule may be any nucleic acid molecule encoding GPR75 Ala110fs, Ala116Thr, Tyr207Cys, Gln234Stop, Arg236fs, or Cys400fs. The GPR75 missense variant nucleic acid molecule may also be Lys404 * It could be any nucleic acid molecule encoding Ser219fs, or it could be c.-110+1G>A.

[0041] GPR75 missense variant nucleic acid molecules include 2:53853134:T:G, 2:53853135:T:G, 2:53853136:A:C, 2:53853200:GGT:G, 2:53853245:GT:G, 2:53853256:G:A, 2:53853352:G:A, 2:53853354:CCA:C, 2:53853382:TG:T, 2:53853502:T:A, 2:53853535:G:A, 2:53853547: T:A, 2:53853560:G:GA, 2:53853641:GTT:G, 2:53853680:CAATTCAAACTGGT:C, 2:53853692:G:T, 2:53853730:G:A, 2:53853 771:G:C, 2:53853853:G:A, 2:53853877:G:A, 2:53853926:G:T, 2:53853927:T:TA, 2:53853946:G:GT, 2:53853967:TGG:T, 2 :53854009:G:A, 2:53854037:A:AG, 2:53854045:ACTTT:A, 2:53854051:T:A, 2:53854057:G:A, 2:53854078:G:A, 2:538540 99:CAG:C, 2:53854135:CAT:C, 2:53854137:TAGAG:T, 2:53854306:G:A, 2:53854380:G:C, 2:53854409:A:AG, 2:53854421:A This includes, but is not limited to, CTACTGG:A, 2:53854474:C:A, 2:53854476:C:CA, 2:53854485:AG:A, 2:53854644:TG:T, 2:53854685:TC:T, 2:53854695:G:T, 2:53854740:TG:T, 2:53854755:A:G, and 2:53859827:C:T (compliant with GRCh38 / hg38 (December 2013) Human Genome Assembly).

[0042] In any of the embodiments described herein, the predicted loss of function polypeptide of GPR75 may be any GPR75 polypeptide having partial loss of function, complete loss of function, predicted partial loss of function, or predicted complete loss of function. In any of the embodiments described herein, the predicted loss of function polypeptide of GPR75 may be any of the GPR75 polypeptides described herein, for example, GPR75 Ala110fs, Ala116Thr, Tyr207Cys, Gln234Stop, Arg236fs, or Cys400fs. The predicted loss of function polypeptide of GPR75 may also be Lys404 * Alternatively, it could be Ser219fs.

[0043] In any of the embodiments described herein, the subject may be obese, or may have an overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or may be consuming excessive amounts of food. In any of the embodiments described herein, the subject may be obese. In any of the embodiments described herein, the subject may be overweight. In any of the embodiments described herein, the subject may have an elevated BMI. In any of the embodiments described herein, the subject may have an elevated body fat mass, body fat percentage, or body fat volume. In any of the embodiments described herein, the subject may be consuming excessive amounts of food.

[0044] Symptoms of obesity include, but are not limited to, excessive accumulation of body fat (especially around the waist), shortness of breath, increased sweating, snoring, inability to cope with sudden physical activity, severe daily fatigue, back and joint pain, and skin problems (due to fluid retention in skin folds).

[0045] This disclosure provides a method for treating a subject having obesity, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0046] This disclosure also provides a method for treating a subject having an overweight body weight, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0047] This disclosure also provides a method for treating subjects with elevated BMI, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0048] This disclosure also provides a method for treating subjects with elevated body fat mass, body fat percentage, or body fat volume, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0049] This disclosure also provides a method for treating a subject who has ingested excessive amounts of food, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0050] This disclosure also provides a method for treating a subject to prevent weight gain or maintain weight loss, the method comprising administering a GPR75 inhibitor to the subject. In some embodiments, the subject is heterozygous for a GPR75 reference or a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0051] In any of the embodiments described herein in which a subject is treated with a GPR75 inhibitor, the subject may be a GPR75 reference (i.e., the subject does not have a copy of the GPR75 missense variant nucleic acid molecule encoding the predicted loss-of-function GPR75 polypeptide). In any of the embodiments described herein in which a subject is treated with a GPR75 inhibitor, the subject may be heterozygous for the GPR75 missense variant nucleic acid molecule encoding the predicted loss-of-function GPR75 polypeptide (i.e., the subject has only a single copy of the reference GPR75 nucleic acid molecule). It is not necessary to determine the subject's genotype at the time of administration of the GPR75 inhibitor.

[0052] In some embodiments, the GPR75 inhibitor comprises an inhibitory nucleic acid molecule. Examples of inhibitory nucleic acid molecules include, but are not limited to, antisense nucleic acid molecules, small interfering RNA (siRNA), and short hairpin RNA (shRNA). Such inhibitory nucleic acid molecules may be designed to target any region of the GPR75 nucleic acid molecule, such as an mRNA molecule. In some embodiments, the antisense RNA, siRNA, or shRNA hybridizes with a sequence within the GPR75 genomic nucleic acid molecule or mRNA molecule to reduce the expression of the GPR75 polypeptide in the target cells. In some embodiments, the GPR75 inhibitor comprises an antisense RNA that hybridizes with the GPR75 genomic nucleic acid molecule or mRNA molecule to reduce the expression of the GPR75 polypeptide in the target cells. In some embodiments, the antisense nucleic acid molecule comprises or consists of the nucleotide sequences shown in SEQ ID NOs: 109-529. In some embodiments, the GPR75 inhibitor comprises an siRNA that hybridizes with the GPR75 genomic nucleic acid molecule or mRNA molecule to reduce the expression of the GPR75 polypeptide in the target cells. In some embodiments, the siRNA molecule comprises or consists of nucleotide sequences (sense and antisense strand pairs) shown in SEQ ID NOs. 530-1457 (i.e., SEQ ID NOs. 530 and 531, respectively, which form the sense and antisense strands of the siRNA molecule; similarly for the sense and antisense strands of the remaining siRNA molecules shown in SEQ ID NOs. 530-1457). In some embodiments, the GPR75 inhibitor comprises shRNA that hybridizes with a GPR75 genomic nucleic acid molecule or mRNA molecule to reduce the expression of the GPR75 polypeptide in the target cell. Suitable GPR75-specific siRNAs and shRNAs are disclosed, for example, in Garcia et al., Circ. Res., 2017, 120, 1776-1788.

[0053] The inhibitory nucleic acid molecules disclosed herein may include RNA, DNA, or both RNA and DNA. Inhibitory nucleic acid molecules may also be ligated or fused with heterologous nucleic acid sequences (e.g., those in a vector) or heterologous labels. For example, the inhibitory nucleic acid molecules disclosed herein may exist in a vector containing the inhibitory nucleic acid molecule and the heterologous nucleic acid sequence, or as an exogenous donor sequence containing them. Inhibitory nucleic acid molecules may also be ligated or fused with heterologous labels. Labels may be directly detectable (e.g., fluorophores) or indirectly detectable (e.g., haptens, enzymes, or fluorophore enchanters). Such labels may be detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Such labels include, for example, radioactive labels, pigments, dyes, chromogens, spin labels, and fluorescent labels. Labels may also be, for example, chemiluminescent substances, metal-containing substances, or enzymes, in the case of enzymes, resulting in enzyme-dependent secondary signal generation. The term “label” can also refer to a “tag” or hapten that can selectively bind to a conjugate molecule, so that the conjugate molecule can be used to generate a detectable signal when later added with a substrate. For example, biotin can be used as a tag with the avidin conjugate or streptavidin conjugate of horseradish peroxidase (HRP), and bound to this tag, and then examined using a chromogenic substrate (e.g., tetramethylbenzidine (TMB)) or a fluorescent substrate to detect the presence of HRP. Exemplary labels that can be used as tags to facilitate purification include, but are not limited to, myc, HA, FLAG, or 3XFLAG, 6XHis, or polyhistidine, glutathione-S-transferase (GST), maltose-binding proteins, epitope tags, or the Fc portion of immunoglobulins. Many labels include, for example, particles, fluorophores, haptens, enzymes and chromogenic substrates for enzymes, as well as chemiluminescent substrates, and other labels.

[0054] The disclosed inhibitory nucleic acid molecules may include nucleotides or unnatural or modified nucleotides, such as nucleotide analogs or substitute nucleotides. Such nucleotides may include nucleotides containing modified bases, sugars, or phosphate groups, or nucleotides in which unnatural moieties are incorporated into their structure. Examples of unnatural nucleotides include, but are not limited to, dideoxynucleotides, biotinylated nucleotides, aminated nucleotides, deaminated nucleotides, alkylated nucleotides, benzylated nucleotides, and fluorophore-labeled nucleotides.

[0055] The inhibitory nucleic acid molecules disclosed herein may also comprise one or more nucleotide analogs or substitutions. A nucleotide analog is a nucleotide containing a modification to either the base moiety, sugar moiety, or phosphate moiety. Modifications to the base moiety include, but are not limited to, natural and synthetic modifications of A, C, G, and T / U, as well as various purine or pyrimidine bases, such as pseudouridine, uracil-5-yl, hypoxanthin-9-yl(I), and 2-aminoadenine-9-yl. Modified bases include 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl derivatives and other alkyl derivatives of adenine and guanine, 2-propyl derivatives and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine, and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azouracil, cytosine and thymine, and 5-uracil (shrubyl). Examples include, but are not limited to, douracil, 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (e.g., 5-bromo), 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine, 7-methyladenine, 8-azaguanine, 8-azaadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, and 3-deazaadenine.

[0056] Nucleotide analogs may also include modifications of the sugar moiety. Modifications to the sugar moiety include, but are not limited to, natural modifications as well as synthetic modifications of ribose and deoxyribose. Sugar modifications include, at the 2' position, the modification of OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl, and alkynyl are substituted or unsubstituted C 1~10 alkyl or C 2~10 alkenyl, and C 2~10 alkynyl, including but not limited to the modifications that may be such. Exemplary 2' sugar modifications also include -O[(CH2) n O] m CH3, -O(CH2) n OCH3, -O(CH2) n NH2, -O(CH2) n CH3, -O(CH2) n -ONH2, and -O(CH2) n ON[(CH2) n CH3)]2 (wherein n and m are independently from 1 to about 10), including but not limited thereto. Other modifications at the 2' position include C 1~10Examples of modified sugars include, but are not limited to, alkyl groups, substituted lower alkyl groups, alkaryl groups, aralkyl groups, O-alkaryl or O-aralkyl groups, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl groups, heterocycloalkaryl groups, aminoalkylamino groups, polyalkylamino groups, substituted silyl groups, RNA cleavage groups, reporter groups, intercalators, groups for improving the pharmacokinetic properties of oligonucleotides, or groups for improving the pharmacodynamic properties of oligonucleotides, and other substituents having similar properties. Similar modifications can also occur at other positions of the sugar, particularly on the 3'-terminal nucleotide or at the 3' position of the sugar in oligonucleotides linked at 2'-5', and at the 5' position of the 5'-terminal nucleotide. Modified sugars may also include those containing modifications at ring-bridged oxygens, such as CH2 and S. Nucleotide sugar analogs may also have sugar mimes, such as cyclobutyl moieties replacing pentofuranosyl sugars.

[0057] Nucleotide analogs can also be modified at the phosphate moiety. Modified phosphate moieties may include, but are not limited to, those that modify the bond between two nucleotides to contain phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotryesters, aminoalkyl phosphotryesters, methyl and other alkylphosphonates (including 3'-alkylene phosphonates and chiral phosphonates), phosphinates, phosphoramidates (including 3'-aminophosphoramidates and aminoalkylphosphoramidates), thionophosphoramidates, thionoalkylphosphoramidates, thionoalkyl phosphotryesters, and boranophosphates. These phosphate or modified phosphate bonds between two nucleotides can be 3'-5' or 2'-5' bonds, and the bonds may contain inverted polarity, such as 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts, and free acid forms may also be included. Peptide nucleic acids (PNAs) can also be used as substitute nucleotides.

[0058] In some embodiments, the antisense nucleic acid molecule is a gapmer, thereby having 2'-methoxyethyl (2'-MOE) modifications in the first 1-7 nucleotides of the 5' and 3' ends, respectively. In some embodiments, the first 5 nucleotides of the 5' and 3' ends each have 2'-MOE modifications. In some embodiments, the first 1-7 nucleotides of the 5' and 3' ends are RNA nucleotides. In some embodiments, the first 5 nucleotides of the 5' and 3' ends are RNA nucleotides. In some embodiments, each of the skeletal bonds between nucleotides is a phosphorothioate bond.

[0059] In some embodiments, the siRNA molecule has terminal modifications. In some embodiments, the 5' end of the antisense chain is phosphorylated. In some embodiments, a hydrolyzable 5'-phosphate analog, such as 5'-(E)-vinyl-phosphonate, is used.

[0060] In some embodiments, the siRNA molecule has skeletal modifications. In some embodiments, modifying phosphodiester groups linking consecutive ribose nucleosides have been shown to enhance the stability and in vivo bioavailability of the siRNA. By replacing the non-ester group (-OH, =O) of the phosphodiester bond with sulfur, boron, or acetate, phosphorothioate bonds, boranophosphate bonds, and phosphonoacetate bonds can be obtained. In addition, substitution of the phosphodiester group with a phosphotryester can promote the uptake of siRNA into cells and retention in serum components by removing the negative charge of the siRNA. In some embodiments, the siRNA molecule has sugar modifications. In some embodiments, the sugar is deprotonated (a reaction catalyzed by exonucleases and endonucleases), so that the 2'-hydroxyl acts as a nucleophile and attacks the adjacent phosphorus within the phosphodiester bond. Such alternatives include 2'-O-methyl, 2'-O-methoxyethyl, and 2'-fluoro modifications.

[0061] In some embodiments, the siRNA molecule has base modification. In some embodiments, the base can be replaced with a modified base such as pseudouridine, 5'-methylcytidine, N6-methyladenosine, inosine, and N7-methylguanosine.

[0062] In some embodiments, siRNA molecules are conjugated with lipids. Conjugating lipids to the 5' or 3' end of siRNA can improve in vivo bioavailability by enabling association between the lipid and serum lipoproteins. Typical lipids include, but are not limited to, cholesterol and vitamin E, as well as fatty acids such as palmitate and tocopherol.

[0063] In some embodiments, a typical siRNA has the following formula. Sense: mN * mN * / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / * mN * / 32FN / Antisense: / 52FN / * / i2FN / * mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN * N * N (Here, "N" is a base, "2F" is a 2'-F modification, "m" is a 2'-O-methyl modification, "I" is an internal base, * (This refers to phosphorothioate skeleton bonding.)

[0064] This disclosure also provides vectors comprising one or more inhibitory nucleic acid molecules disclosed herein. In some embodiments, the vector comprises one or more inhibitory nucleic acid molecules disclosed herein and heterogeneous nucleic acids. The vector may be a viral vector or a non-viral vector capable of transporting nucleic acid molecules. In some embodiments, the vector is a plasmid or cosmid (e.g., a circular double-stranded DNA in which additional DNA segments can be ligated). In some embodiments, the vector is a viral vector in which additional DNA segments can be ligated into a viral genome. Examples of expression vectors include, but are not limited to, plasmids, cosmids, retroviruses, adenoviruses, adeno-associated viruses (AAVs), plant viruses (e.g., cauliflower mosaic virus and tobacco mosaic virus), yeast artificial chromosomes (YACs), Epstein-Barr (EBV)-derived episomes, and other expression vectors known in the art.

[0065] This disclosure also provides compositions comprising one or more of the inhibitory nucleic acid molecules disclosed herein. In some embodiments, the compositions are pharmaceutical compositions. In some embodiments, the compositions comprise a carrier and / or excipients. Examples of carriers include, but are not limited to, poly(lactic acid) (PLA) microspheres, D,L-lactic acid-glycolic acid copolymer (PLGA) microspheres, liposomes, micelles, reverse micelles, lipid cocreates, and lipid microtubules. The carrier may comprise a buffer salt solution such as PBS or HBSS.

[0066] In some embodiments, a GPR75 inhibitor comprises a nuclease agent that induces one or more nicks or double-strand breaks in a recognition sequence(s), or a DNA-binding protein that binds to a recognition sequence within the GPR75 genomic nucleic acid molecule. The recognition sequence may be located within the coding region of the GPR75 gene or within a regulatory region that affects gene expression. The recognition sequence of the DNA-binding protein or nuclease agent may be located in an intron, exon, promoter, enhancer, regulatory region, or any non-protein-coding region. The recognition sequence may contain or be adjacent to the start codon of the GPR75 gene. For example, the recognition sequence may be located about 10, 20, 30, 40, 50, 100, 200, 300, 400, 500, or 1,000 nucleotides from the start codon. As another example, two or more nuclease agents can be used, each targeting a nuclease recognition sequence that contains or is adjacent to a start codon. As another example, two nucleases can be used, one targeting a nuclease recognition sequence containing or adjacent to a start codon, and the other targeting a nuclease recognition sequence containing or adjacent to a stop codon, such that cleavage by these nucleases can delete the coding region between the two nuclease recognition sequences. Any nuclease that induces nicks or double-strand breaks in a desired recognition sequence can be used in the methods and compositions disclosed herein. Any DNA-binding protein that binds to a desired recognition sequence can be used in the methods and compositions disclosed herein.

[0067] Suitable nucleases and DNA-binding proteins for use herein include, but are not limited to, zinc finger proteins or zinc finger nuclease (ZFN) pairs, activator-like effector (TALE) proteins or activator-like effector nucleases (TALENs), or clustered, regularly dispersed short palindromic repeats (CRISPR) / CRISPR-related (Cas) systems. Recognition sequences can vary in length, for example, approximately 30–36 bp for zinc finger proteins or ZFN pairs, approximately 15–18 bp for each ZFN, approximately 36 bp for TALE proteins or TALENs, and approximately 20 bp for CRISPR / Cas guide RNAs.

[0068] In some embodiments, the CRISPR / Cas system can be used to modify intracellular GPR75 genomic nucleic acid molecules. The methods and compositions disclosed herein can utilize the CRISPR-Cas system by utilizing a CRISPR complex (containing a guide RNA (gRNA) complexed with a Cas protein) for site-specific cleavage of the GPR75 nucleic acid molecule.

[0069] Cas proteins generally contain at least one RNA recognition domain or RNA binding domain capable of interacting with gRNA. Cas proteins may also contain nuclease domains (e.g., DNase or RNase domains), DNA binding domains, helicase domains, protein-protein interaction domains, dimerization domains, and other domains. Suitable Cas proteins include, for example, wild-type Cas9 and wild-type Cpf1 proteins (e.g., FnCpf1). Cas proteins may have complete cleavage activity to produce double-strand breaks in GPR75 genomic nucleic acid molecules, or they may be nicasses that produce single-strand breaks in GPR75 genomic nucleic acid molecules. Further examples of Cas proteins include Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas5e (CasD), Cas6, Cas6e, Cas6f, Cas7, Cas8a1, Cas8a2, Cas8b, Cas8c, Cas9 (Csn1 or Csx12), Cas10, Cas10d, CasF, CasG, CasH, Csy1, Csy2, Csy3, Cse1 (CasA), Cse2 (CasB), Cse3 (CasE), C Examples include, but are not limited to, se4(CasC), Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, and Cu1966, as well as their homologs or modified forms. Cas proteins can also be operably ligated to heterologous polypeptides as fusion proteins. For example, a Cas protein can be fused to a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Cas proteins can be provided in any form. For example, Cas proteins can be provided in the form of Cas proteins complexed with other proteins, such as gRNA.Alternatively, the Cas protein can be provided in the form of a nucleic acid molecule that codes for the Cas protein, such as RNA or DNA.

[0070] In some embodiments, the gene modifications targeted by the GPR75 genomic nucleic acid molecule can be brought into contact with a Cas protein and one or more gRNAs that hybridize with one or more gRNA recognition sequences within the target genomic locus of the GPR75 genomic nucleic acid molecule. For example, the gRNA recognition sequence may be located within the region of SEQ ID NO: 1. The gRNA recognition sequence may also include, or be adjacent to, positions corresponding to positions 5,540-5,546, 5,557, 5,911, 5,920-5,923, or 6,411 as described in SEQ ID NO: 1. For example, the gRNA recognition sequence may be located approximately 1000, 500, 400, 300, 200, 100, 50, 45, 40, 35, 30, 25, 20, 100, 15, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, or 5 nucleotides from the positions corresponding to positions 5,540–5,546, 5,557, 5,911, 5,920–5,923, or 6,411 as described in Sequence ID No. 1. The gRNA recognition sequence may contain or be close to the start codon or stop codon of the GPR75 genomic nucleic acid molecule. For example, the gRNA recognition sequence may be located approximately 10, 20, 30, 40, 50, 100, 200, 300, 400, 500, or 1000 nucleotides from the start or stop codon.

[0071] The gRNA recognition sequence within the target genomic locus of the GPR75 genomic nucleic acid molecule is located near a protospacer adjacency motif (PAM) sequence, which is a 2-6 base pair DNA sequence immediately following the DNA sequence targeted by the Cas9 nuclease. The canonical PAM is the sequence 5'-NGG-3', where "N" is any nucleic acid base followed by two guanine ("G") nucleic acid bases. While the gRNA can transport Cas9 anywhere in the genome for gene editing, editing cannot occur anywhere other than the site where Cas9 recognizes the PAM. In addition, 5'-NGA-3' can be a highly efficient non-canonical PAM for human cells. Generally, the PAM is approximately 2-6 nucleotides downstream of the DNA sequence targeted by the gRNA. The PAM can be adjacent to the gRNA recognition sequence. In some embodiments, the gRNA recognition sequence may be adjacent to the PAM at its 3' end. In some embodiments, the gRNA recognition sequence may be adjacent to the PAM at its 5' end. For example, the cleavage site of the Cas protein can be approximately 1 to 10, 2 to 5, or 3 base pairs upstream or downstream of the PAM sequence. In some embodiments (e.g., when using Cas9 or a closely related Cas9 derived from S. pyogenes), the non-complementary strand PAM sequence may be 5'-NGG-3' (where N is any DNA nucleotide) and is the 3' end immediately adjacent to the gRNA recognition sequence on the non-complementary strand of the target DNA. Thus, the complementary strand PAM sequence may be 5'-CCN-3' (where N is any DNA nucleotide) and is the 5' end immediately adjacent to the gRNA recognition sequence on the complementary strand of the target DNA.

[0072] gRNA is an RNA molecule that binds to the Cas protein and targets the Cas protein to specific locations within the GPR75 genomic nucleic acid molecule. An exemplary gRNA is a gRNA effective in inducing the Cas enzyme to bind to and cleave the GPR75 genomic nucleic acid molecule, and the gRNA contains a DNA targeting segment that hybridizes with a gRNA recognition sequence within the GPR75 genomic nucleic acid molecule that includes or is adjacent to the positions corresponding to positions 5,540–5,546, 5,557, 5,911, 5,920–5,923, or 6,411 as described in Sequence ID No. 1. For example, a gRNA may be selected to hybridize with a gRNA recognition sequence located approximately 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 300, 400, 500, or 1,000 nucleotides from the positions corresponding to positions 5,540–5,546, 5,557, 5,911, 5,920–5,923, or 6,411 as described in SEQ ID NO: 1. Other exemplary gRNAs include DNA targeting segments that hybridize with gRNA recognition sequences located within the GPR75 genomic nucleic acid molecule, either containing or adjacent to a start or stop codon. For example, the gRNA may be selected to hybridize with a gRNA recognition sequence located approximately 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 400, 500, or 1,000 nucleotides from the start codon, or with a gRNA recognition sequence located approximately 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 400, 500, or 1,000 nucleotides from the stop codon. A suitable gRNA may contain approximately 17–25 nucleotides, approximately 17–23 nucleotides, approximately 18–22 nucleotides, or approximately 19–21 nucleotides. In some embodiments, the gRNA may contain 20 nucleotides.

[0073] Examples of suitable gRNA recognition sequences located within the human GPR75 reference gene are shown in Table 1 as sequence numbers 61-98.

[0074] [Table 1]

[0075] The Cas protein and gRNA form a complex, and the Cas protein cleaves the target GPR75 genomic nucleic acid molecule. The Cas protein can cleave the nucleic acid molecule at a site inside or outside the nucleic acid sequence present on the target GPR75 genomic nucleic acid molecule to which the gRNA's DNA targeting segment binds. For example, the formation of a CRISPR complex (containing gRNA that hybridizes with a gRNA recognition sequence and complexes with the Cas protein) can result in cleavage of one or both strands within or near the nucleic acid sequence present on the GPR75 genomic nucleic acid molecule to which the gRNA's DNA targeting segment binds (e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50 or more base pairs from that nucleic acid sequence).

[0076] Such methods may result in a GPR75 genomic nucleic acid molecule in which, for example, the region of sequence number 1 is disrupted, the start codon is disrupted, the stop codon is disrupted, or the coding sequence is disrupted or deleted. Optionally, the cell can be further contacted with one or more additional gRNAs that hybridize with additional gRNA recognition sequences within the target genomic locus of the GPR75 genomic nucleic acid molecule. By contacting the cell with one or more additional gRNAs (e.g., a second gRNA that hybridizes with a second gRNA recognition sequence), cleavage by the Cas protein can create two or more double-strand breaks or two or more single-strand breaks.

[0077] In some embodiments, the GPR75 inhibitor comprises a small molecule. In some embodiments, the GPR75 inhibitor is an inhibitor of CCL5(RANTES), but is not limited to [ 44 AANA 47This includes ]-RANTES and Met-CCL5 (see Braunersreuther et al., Arteriosclerosis, Thrombosis, and Vasc. Biol., 2008, 28, 1090-1096, Proudfoot et al., J. Biol. Chem., 1999, 274, 32478-32485, and Matsui et al., J. Neroimmunol., 2002, 128, 16-22). In some embodiments, GPR75 inhibitors are inhibitors of the interaction between GPR75 and 20-hydroxyeicosatetraenoic acid (20-HETE), and include, but are not limited to, fatty acid analogs, terminal acetylene fatty acids, terminal dibromo fatty acids, sulfonated fatty acids, TS-011, Het0016, 5,14,20-HEDE, 5,14,20-HEDGE, 6,15,20-HEDE, 17-ODYA, DDMS, DDBB, 2,5,8,11,14,17-hexaoxanadecan-19-yl-20-hydroxyeicosa-6(z),15(z)-dienoate (20-sola), and 6(z),15(z)-hydroxyeicosa-6,15-dienamide-dienoic acid (aaa) (Miyata et al. See also al., Br.J.Pharmacol.,2001,133,325-329, Miyata et al.,J.Pharmacol.Exp.Ther.,2005,314,77-85, Pandey et al.,J.Pharmacol.Exp.Ther.,2017,363,412-418, and Savas et al.,J.Biol.Chem.,2016,291,16904-16919). In some embodiments, the GPR75 inhibitor is one of the antagonists described in PCT Publication WO2017 / 156164. In some embodiments, the GPR75 inhibitor is an inhibitor of the interaction between GPR75 and 20-HETE, and is a blocking antibody.

[0078] In some embodiments, the therapeutic method further includes detecting the absence of a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide in a biological sample from a subject. As used throughout this disclosure, “GPR75 missense variant nucleic acid molecule” is any GPR75 nucleic acid molecule (e.g., a genomic nucleic acid molecule, an mRNA molecule, or a cDNA molecule) encoding a GPR75 polypeptide having partial loss-of-function, complete loss-of-function, predicted partial loss-of-function, or predicted complete loss-of-function.

[0079] This disclosure also provides a method for treating a subject with a therapeutic agent for treating or suppressing obesity, wherein the subject is obese. In some embodiments, the method includes determining whether the subject has a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide, which is done by obtaining or having obtained a biological sample from the subject and performing or having performed a sequence analysis on the biological sample to determine whether the subject has a genotype containing the GPR75 missense variant nucleic acid molecule. If the subject is GPR75 reference, the therapeutic agent for treating or suppressing obesity and / or the GPR75 inhibitor is administered to or continues to be administered to the subject at a standard dose. If the subject is heterozygous for the GPR75 missense variant nucleic acid molecule, the therapeutic agent for treating or suppressing obesity and / or the GPR75 inhibitor is administered to or continues to be administered to the subject at the same or a lower dose than the standard dose. The presence of a genotype containing a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide indicates that the subject has a reduced risk of developing obesity. In some embodiments, the subject is a GPR75 reference. In some embodiments, the subject is heterozygous for a GPR75 missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide.

[0080] For subjects that have been genotyped or determined to be heterozygous for either GPR75 reference or GPR75 missense variant nucleic acid molecules, such subjects may be treated with a GPR75 inhibitor as described herein.

[0081] The presence or absence of GPR75 missense variant nucleic acid molecules in a biological sample from a subject, and / or the presence or absence of GPR75 missense variant nucleic acid molecules in a subject, can be detected by any of the methods described herein. In some embodiments, these methods can be performed in vitro. In some embodiments, these methods can be performed in situ. In some embodiments, these methods can be performed in vivo. In any of these embodiments, the nucleic acid molecules may be present in cells obtained from the subject.

[0082] In some embodiments, if the subject is a GPR75 reference, the subject is also administered a therapeutic agent and / or GPR75 inhibitor for treating or suppressing obesity at a standard dose. In some embodiments, if the subject is heterozygous for a GPR75 missense variant nucleic acid molecule, the subject is also administered a therapeutic agent and / or GPR75 inhibitor for treating or suppressing obesity at the same or a lower dose than the standard dose.

[0083] In some embodiments, the treatment method further includes detecting the absence of a predicted loss-of-function polypeptide of GPR75 in a biological sample from the subject. In some embodiments, if the subject does not have a predicted loss-of-function polypeptide of GPR75, the subject is also administered a therapeutic agent and / or GPR75 inhibitor for treating or suppressing obesity at a standard dose. In some embodiments, if the subject does have a predicted loss-of-function polypeptide of GPR75, the subject is also administered a therapeutic agent and / or GPR75 inhibitor for treating or suppressing obesity at the same or a lower dose than the standard dose.

[0084] This disclosure also provides a method for treating a subject with a therapeutic agent for treating or suppressing obesity, wherein the subject is obese. In some embodiments, the method includes determining whether the subject has a predicted loss-of-function polypeptide of GPR75, which is done by obtaining or having obtained a biological sample from the subject and performing or having performed an assay on the biological sample to determine whether the subject has a predicted loss-of-function polypeptide of GPR75. If the subject does not have a predicted loss-of-function polypeptide of GPR75, the therapeutic agent for treating or suppressing obesity and / or a GPR75 inhibitor is administered to or continues to be administered to the subject at a standard dose. If the subject has a predicted loss-of-function polypeptide of GPR75, the therapeutic agent for treating or suppressing obesity and / or a GPR75 inhibitor is administered to or continues to be administered to the subject at the same or a lower dose than the standard dose. The presence of a predicted loss-of-function polypeptide of GPR75 indicates that the subject has a reduced risk of developing obesity. In some embodiments, the subject has a predicted loss-of-function polypeptide of GPR75. In some embodiments, the subject does not have the predicted loss-of-function polypeptide of GPR75.

[0085] The presence or absence of the predicted loss-of-function polypeptide of GPR75 in a biological sample from a subject, and / or the determination of whether the subject has the predicted loss-of-function polypeptide of GPR75, can be performed by any of the methods described herein. In some embodiments, these methods can be performed in vitro. In some embodiments, these methods can be performed in situ. In some embodiments, these methods can be performed in vivo. In any of these embodiments, the polypeptide may be present in cells obtained from the subject.

[0086] Examples of therapeutic agents for treating or suppressing obesity and / or BMI increase include, but are not limited to, sibutramine, orlistat, phentermine, lorcaserin, naltrexone, liraglutide, diethylpropion, bupropion, metformin, pramulintide, topiramate, and zonisamide, or any combination thereof. Furthermore, examples of therapeutic agents for treating or suppressing obesity and / or BMI increase include, but are not limited to, sibutramine, orlistat, phentermine and metformin, lorcaserin, bupropion and naltrexone, liraglutide, phentermine and diethylpropion, bupropion, metformin, pramulintide, topiramate, and zonisamide, or any combination thereof.

[0087] In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is a GLP1R agonist, including but not limited to BYETTA® or BYDUREON® (exenatide), VICTOZA® or SAXENDA® (liraglutide), LYXUMIA® or ADLYXIN® (lixisenatide), TANZEUM® (albiglutide), TRULICITY® (dulaglutide), and OZEMPIC® (semaglutide), or any combination thereof. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is BYETTA® (exenatide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is BYETTA® (exenatide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is VICTOZA® (liraglutide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is SAXENDA® (liraglutide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is LYXUMIA® (lixisenatide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is ADLYXIN® (lixisenatide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is TANZEUM® (albiglutide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is TRULICITY® (dulaglutide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is OZEMPIC® (semaglutide). In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is selected from exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, and semaglutide, or any combination thereof.In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is exenatide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is liraglutide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is lixisenatide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is albiglutide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is dulaglutide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or BMI increase is semaglutide.

[0088] In some embodiments, the therapeutic agent for treating or suppressing obesity and / or reducing BMI is a melanocortin 4 receptor (MC4R) agonist. In some embodiments, the MC4R agonist comprises a protein, peptide, nucleic acid molecule, or small molecule. In some embodiments, the protein is a peptide analog of MC4R. In some embodiments, the peptide is cetomelanotide. In some embodiments, the therapeutic agent for treating or suppressing obesity and / or reducing BMI is a combination of cetomelanotide with one or more of sibutramine, orlistat, phentermine, lorcaserin, naltrexone, liraglutide, diethylpropion, bupropion, metformin, pramlintide, topiramate, and zonisamide. In some embodiments, the MC4R agonist is a peptide containing the amino acid sequence His-Phe-Arg-Trp. In some embodiments, the small molecule is 1,2,3R,4-tetrahydroisoquinoline-3-carboxylic acid. In some embodiments, the MC4R agonist is ALB-127158(a).

[0089] In some embodiments, the dose of an obesity-treating or deterrent agent and / or GPR75 inhibitor can be reduced by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% (i.e., less than the standard dose) for subjects heterozygous for the GPR75 missense variant nucleic acid molecule compared to subjects who are GPR75 reference (and may receive a standard dose). In some embodiments, the dose of an obesity-treating or deterrent agent and / or GPR75 inhibitor can be reduced by approximately 10%, 20%, 30%, 40%, or 50%. In addition, the dose of an obesity-treating or deterrent agent and / or GPR75 inhibitor can be administered at a lower frequency for subjects heterozygous for the GPR75 missense variant nucleic acid molecule compared to subjects who are GPR75 reference.

[0090] Therapeutic agents and / or GPR75 inhibitors that treat or suppress obesity may be repeated, for example, after 1 day, 2 days, 3 days, 5 days, 1 week, 2 weeks, 3 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 2 months, or 3 months. Repeated doses may be the same or different doses. Administration may be repeated once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, or more. For example, according to a particular dosage regimen, subjects may be treated over a long period, such as six months, one year, or longer.

[0091] Therapeutic agents and / or GPR75 inhibitors for treating or suppressing obesity may be administered by any preferred route, including but not limited to parenteral, intravenous, oral, subcutaneous, intra-arterial, intracranial, intrathecal, intraperitoneal, topical, intranasal, or intramuscular. Pharmaceutical compositions for administration should preferably be sterile, substantially isotonic, and manufactured under GMP conditions. Pharmaceutical compositions may be provided in unit dosage forms (i.e., single-dose doses). Pharmaceutical compositions may be formulated using one or more physiologically and pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. The formulation depends on the chosen route of administration. The term "pharmaceutically acceptable" means that the carrier, diluent, excipient, or adjuvant is compatible with the other components of the formulation and is not substantially harmful to its recipient.

[0092] In some embodiments, therapeutic agents and / or GPR75 inhibitors (e.g., any of the inhibitory nucleic acid molecules disclosed herein) that treat or suppress obesity are administered intrathecally (i.e., by introducing the therapeutic agent into the subarachnoid space or spinal canal of the spinal cord, or into anatomical or latent spaces within the sheath (including, in non-limiting examples, the arachnoid membrane of the brain or spinal cord) so that the therapeutic agent can reach the cerebrospinal fluid of the target). In some embodiments, intrathecal administration causes the therapeutic agent to act on the cortex, cerebellum, striatum, cervical vertebrae, lumbar vertebrae, or thoracic vertebrae, but are not limited to these. The intrathecally administered therapeutic agent can ultimately act on targets throughout the central nervous system. In some embodiments, intrathecal administration is into the cisterna magna or near the lumbar region or area. In some embodiments, intrathecal administration to the lumbar region or area delivers the therapeutic agent to the distal spinal canal. Exemplary methods of intrathecal administration are described, for example, in Lazorthes et al., Advances in Drug Delivery Systems and Applications in Neurosurgery, 143-192. In some embodiments, intrathecal administration is by injection, bolus injection, catheter, or pump. In some embodiments, intrathecal administration is by lumbar puncture. In some embodiments, the pump is an osmotic pump. In some embodiments, the pump is implanted in the subarachnoid space of the spinal canal, under the skin of the abdomen, or posteriorly in the chest wall. In some embodiments, intrathecal administration is by an intrathecal delivery system of a therapeutic substance, comprising a reservoir containing a certain amount of the therapeutic substance and a pump configured to deliver at least a portion of the therapeutic substance contained in the reservoir. In some embodiments, intrathecal administration is via intermittent or continuous access to an implantable intrathecal drug delivery device (IDDD). In some embodiments, the therapeutic substance is an inhibitory nucleic acid molecule. In some embodiments, the amount of nucleic acid molecules administered intrathecally is in the range of about 10 μg to about 2 mg, about 50 μg to about 1500 μg, or about 100 μg to about 1000 μg. In some embodiments, the therapeutic agent is placed within the pharmaceutical composition. In some embodiments, the pharmaceutical composition does not contain preservatives.

[0093] As used herein, the terms “to treat,” “to treat,” and “treatment,” as well as “to prevent,” “to prevent,” and “prevent,” respectively, refer to eliciting a desired biological response, such as a therapeutic effect and a preventive effect. In some embodiments, a therapeutic effect includes one or more of the following after administration of a drug or a composition containing a drug: a reduction in obesity, a decrease in the severity of obesity (e.g., a reduction or suppression of the onset of obesity), a reduction in symptoms and obesity-related effects, a delay in the onset of symptoms and obesity-related effects, a reduction in the severity of symptoms of obesity-related effects, a reduction in the severity of acute episodes, a reduction in the number of symptoms and obesity-related effects, a shortening of the incubation period for symptoms and obesity-related effects, an improvement in symptoms and obesity-related effects, a reduction in secondary symptoms, a reduction in secondary infections, prevention of obesity relapses, a decrease in the number or frequency of recurrent episodes, a prolongation of the incubation period between symptomatic episodes, a prolongation of the time to sustained progression, promotion of remission, induction of remission, increased remission, accelerated recovery, or increased effectiveness of alternative treatments or decreased resistance thereto, and / or an extension of the lifespan of the affected host animal. Preventive effects may include complete or partial avoidance / suppression or delay of the onset / progression of obesity after administration of the treatment protocol (e.g., complete or partial avoidance / suppression or delay), or extension of the survival period of affected host animals. Treatment of obesity includes treatment of subjects already diagnosed with any form of obesity at any clinical stage or clinical symptom, delaying the onset or progression or exacerbation or worsening of symptoms or signs of obesity, and / or prevention and / or reduction of the severity of obesity.

[0094] This disclosure also provides a method for identifying subjects at high risk of developing obesity, overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake. In some embodiments, the method includes determining, or having determined, the presence or absence of a GPR75 missense variant nucleic acid molecule (such as a genomic nucleic acid molecule, mRNA molecule, and / or cDNA molecule) encoding a predicted loss-of-function GPR75 polypeptide in a biological sample obtained from a subject. If a subject lacks the GPR75 missense variant nucleic acid molecule (i.e., the subject is classified as GPR75 reference in genotyping), the subject is at high risk of developing obesity. If a subject has the GPR75 missense variant nucleic acid molecule (i.e., the subject is heterozygous or homozygous for the GPR75 missense variant nucleic acid molecule), the subject is at low risk of developing obesity.

[0095] Subjects possessing a single copy of the GPR75 missense variant nucleic acid molecule are protected from developing obesity more effectively than those not possessing a copy of the GPR75 missense variant nucleic acid molecule. While not intended to be limited to any specific theory or mechanism of action, it is believed that subjects are protected from developing obesity by possessing a single copy of the GPR75 missense variant nucleic acid molecule (i.e., heterozygous for the GPR75 missense variant nucleic acid molecule), and that subjects possessing two copies of the GPR75 missense variant nucleic acid molecule (i.e., homozygous for the GPR75 missense variant nucleic acid molecule) may provide further protection compared to subjects with a single copy. Therefore, in some embodiments, a single copy of the GPR75 missense variant nucleic acid molecule may not completely protect subjects from developing obesity, but rather provide partial or incomplete protection. While we do not wish to be bound by any particular theory, there may still be additional factors or molecules involved in the development of obesity that are present in subjects with a single copy of the GPR75 missense variant nucleic acid molecule, and therefore, complete protection against the development of obesity may not be achieved.

[0096] Determining whether a biological sample from a subject contains a GPR75 missense variant nucleic acid molecule, and / or whether a subject contains a GPR75 missense variant nucleic acid molecule, can be done by any of the methods described herein. In some embodiments, these methods can be performed in vitro. In some embodiments, these methods can be performed in situ. In some embodiments, these methods can be performed in vivo. In any of these embodiments, the nucleic acid molecule may be present in cells obtained from the subject.

[0097] In some embodiments, if a subject is identified as being at high risk of developing obesity, the subject is further treated with a therapeutic agent and / or GPR75 inhibitor that treats or suppresses obesity, as described herein. For example, if a subject is a GPR75 reference and therefore at high risk of developing obesity, the subject is administered a GPR75 inhibitor. In some embodiments, such a subject is also administered a therapeutic agent that treats or suppresses obesity. In some embodiments, if a subject is heterozygous for a GPR75 missense variant nucleic acid molecule, the subject is administered a therapeutic agent and / or GPR75 inhibitor that treats or suppresses obesity at the same or lower dose as the standard dose. In some embodiments, the subject is a GPR75 reference. In some embodiments, the subject is heterozygous for a GPR75 missense variant nucleic acid molecule.

[0098] This disclosure also provides a method for detecting the presence or absence of GPR75 missense genomic variant nucleic acid molecules, and / or GPR75 missense variant mRNA molecules, and / or GPR75 missense variant cDNA molecules generated from mRNA molecules in a biological sample from a subject. It should be understood that gene sequences within a population, and the mRNA molecules encoded by such genes, can differ due to polymorphisms such as single nucleotide polymorphisms. The sequences provided herein for GPR75 variant genomic nucleic acid molecules, GPR75 variant mRNA molecules, and GPR75 variant cDNA molecules are illustrative sequences only. Other sequences are also possible for GPR75 variant genomic nucleic acid molecules, variant mRNA molecules, and variant cDNA molecules.

[0099] Biological samples may be derived from any cells, tissues, or bodily fluids from a subject. Biological samples may include any clinically relevant tissue, such as bone marrow samples, tumor biopsy specimens, fine-needle aspiration specimens, or bodily fluid samples (e.g., blood, gingival crevicular fluid, plasma, serum, lymph, ascites, cystic fluid, or urine). In some cases, the sample may include an oral swab. The biological samples used in the methods disclosed herein may vary based on the assay format, the nature of the detection method, and the tissue, cells, or extract used as the sample. Biological samples may be subjected to different treatments depending on the assay employed. For example, when detecting any GPR75 variant nucleic acid molecule, pretreatment designed to isolate or concentrate the biological sample for genomic DNA may be employed. Various techniques can be used for this purpose. When detecting the level of any GPR75 variant mRNA molecule, various techniques can be used to concentrate the biological sample containing the mRNA molecule. Various methods can be used to detect the presence or level of mRNA molecules, or the presence of a specific variant genomic DNA locus.

[0100] In some embodiments, detecting human GPR75 missense variant nucleic acid molecules in a subject involves assaying or genotyping a biological sample obtained from the subject to determine whether the GPR75 genomic nucleic acid molecule in the biological sample, and / or the GPR75 mRNA molecule in the biological sample, and / or the GPR75 cDNA molecule generated from the mRNA molecule in the biological sample, contains one or more mutations that cause or are predicted to cause loss of function (partial or complete).

[0101] In some embodiments, a method for detecting the presence or absence of GPR75 missense variant nucleic acid molecules (e.g., genomic nucleic acid molecules, mRNA molecules, and / or cDNA molecules generated from mRNA) in a subject includes performing an assay on a biological sample obtained from the subject. The assay determines whether the nucleic acid molecules in the biological sample contain a specific nucleotide sequence.

[0102] In some embodiments, the nucleotide sequence lacks the CCAGUAG heptanucleotide at the position corresponding to positions 5,540-5,546 described in SEQ ID NO: 1 (in the case of a genomic nucleic acid molecule); lacks the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 described in SEQ ID NO: 7; lacks the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 described in SEQ ID NO: 8; lacks the CCAGUAG heptanucleotide at the position corresponding to positions 361-367 described in SEQ ID NO: 9; or lacks the CCAGUAG heptanucleotide at positions 600-606 described in SEQ ID NO: 10. The molecule lacks the CCAGUAG heptanucleotide at the corresponding position (in the case of mRNA); the molecule lacks the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 31; the molecule lacks the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32; the molecule lacks the CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33; or the molecule lacks the CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34 (in the case of cDNA).

[0103] In some embodiments, the nucleotide sequence includes adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3 (for genomic nucleic acid molecules); adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 27 (for mRNA molecules); adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 51 (for cDNA molecules).

[0104] In some embodiments, the nucleotide sequence includes thymine at the position corresponding to position 5,911 as described in SEQ ID NO: 4 (for genomic nucleic acid molecules); uracil at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil at the position corresponding to position 971 as described in SEQ ID NO: 28 (for mRNA molecules); thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine at the position corresponding to position 971 as described in SEQ ID NO: 52 (for cDNA molecules).

[0105] In some embodiments, the nucleotide sequence lacks an AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1 (in the case of a genomic nucleic acid molecule); lacks an AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, lacks an AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, lacks an AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or lacks an AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10 (in the case of an mRNA molecule); lacks an AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks an AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, lacks an AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or lacks an AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 34 (in the case of a cDNA molecule).

[0106] In some embodiments, the nucleotide sequence includes a thymine insertion at position 6,411 as described in SEQ ID NO: 6 (for genomic nucleic acid molecules); a uracil insertion at position 1,410 as described in SEQ ID NO: 15, a uracil insertion at position 1,311 as described in SEQ ID NO: 20, a uracil insertion at position 1,232 as described in SEQ ID NO: 25, or a uracil insertion at position 1,471 as described in SEQ ID NO: 30 (for mRNA molecules); a thymine insertion at position 1,410 as described in SEQ ID NO: 39, a thymine insertion at position 1,311 as described in SEQ ID NO: 44, a thymine insertion at position 1,232 as described in SEQ ID NO: 49, or a thymine insertion at position 1,471 as described in SEQ ID NO: 54 (for cDNA molecules).

[0107] In some embodiments, the nucleotide sequence includes guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99 (in the case of a genomic nucleic acid molecule); guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103 (in the case of an mRNA molecule); guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107 (in the case of a cDNA molecule).

[0108] In some embodiments, the nucleotide sequence lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, contains adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, contains thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1, contains an insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, or contains guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99.

[0109] In some embodiments, the nucleotide sequence lacks a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, or at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, or at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or at the position corresponding to positions 600-606 as described in SEQ ID NO: 10. It lacks ochide or its complement, contains adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, contains adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, contains adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22, contains adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, contains uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, or corresponds to position 811 as described in SEQ ID NO: 18 It contains uracil or its complement at the corresponding position, or contains uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, or contains uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, or lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7, or lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 8, or corresponds to positions 741-744 as described in SEQ ID NO: 9 It lacks the AAAG tetranucleotide or its complement at the position, or it lacks the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 10, or it includes an insertion of uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15, or it includes an insertion of uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20, or it includes an insertion of uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, or it includes the 1,It includes an insertion of uracil or its complement at position 471, or guanine or its complement at position 830 as described in SEQ ID NO: 100, or guanine or its complement at position 731 as described in SEQ ID NO: 101, or guanine or its complement at position 652 as described in SEQ ID NO: 102, or guanine or its complement at position 891 as described in SEQ ID NO: 103.

[0110] In some embodiments, the nucleotide sequence lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, or a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, or a CCAGTAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or a CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34. It lacks a nucleotide or its complement, contains adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, contains adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, contains adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, contains adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, contains thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, or the 811 position as described in SEQ ID NO: 42 It contains thymine or its complement at the corresponding position, or contains thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, or contains thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, or lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, or lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, or corresponds to positions 741-744 as described in SEQ ID NO: 33 It lacks the corresponding AAAG tetranucleotide or its complement at the corresponding position, or it lacks the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, or it includes an insertion of thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, or it includes an insertion of thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, or it includes an insertion of thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or it includes the 1,It includes an insertion of thymine or its complement at position 471, or guanine or its complement at position 830 as described in SEQ ID NO: 104, or guanine or its complement at position 731 as described in SEQ ID NO: 105, or guanine or its complement at position 652 as described in SEQ ID NO: 106, or guanine or its complement at position 891 as described in SEQ ID NO: 107.

[0111] In some embodiments, the biological sample comprises cells or cell lysates. Such a method may further include, for example, obtaining a biological sample from a subject that contains a GPR75 genomic nucleic acid molecule or mRNA molecule, and, if mRNA is present, optionally reverse transcribing the mRNA into cDNA. Such an assay may include, for example, determining the identification of their positions in a particular GPR75 nucleic acid molecule. In some embodiments, the method is an in vitro method.

[0112] In some embodiments, the determination step, detection step, or sequence analysis includes sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule, a GPR75 mRNA molecule, or a GPR75 cDNA molecule in a biological sample, wherein the sequenced portion contains one or more mutations that cause, or are predicted to cause, loss of function (partial or complete).

[0113] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes a position corresponding to or its complement at positions 5,540-5,546 as described in SEQ ID NO: 2; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes a position corresponding to or its complement at positions 539-545 as described in SEQ ID NO: 11, a position corresponding to or its complement at positions 440-446 as described in SEQ ID NO: 16, a position corresponding to or its complement at positions 361-367 as described in SEQ ID NO: 21, or a position corresponding to or its complement at positions 600-606 as described in SEQ ID NO: 26; and / or iii) generating GPR75 from mRNA in a biological sample. The procedure involves sequencing at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement to positions 539-545 as described in SEQ ID NO: 35, the position corresponding to or its complement to positions 440-446 as described in SEQ ID NO: 40, the position corresponding to or its complement to positions 361-367 as described in SEQ ID NO: 45, or the position corresponding to or its complement to positions 600-606 as described in SEQ ID NO: 50.The sequenced portion of the GPR75 nucleic acid molecule in the biological sample is determined to be: i) a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, ii) a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or a deletion of the CCAGUAG at the position corresponding to positions 600-606 as described in SEQ ID NO: 10. If the GPR75 nucleic acid molecule in the biological sample contains a heptanucleotide deletion, or iii) a CCAGTAG heptanucleotide deletion at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, a CCAGTAG heptanucleotide deletion at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, a CCAGTAG heptanucleotide deletion at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or a CCAGTAG heptanucleotide deletion at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0114] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 5,557 as described in SEQ ID NO: 3; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 556 as described in SEQ ID NO: 12, the position corresponding to or its complement to position 457 as described in SEQ ID NO: 17, the position corresponding to or its complement to position 378 as described in SEQ ID NO: 22, or the position corresponding to or its complement to position 617 as described in SEQ ID NO: 27; and / or iii) generating GPR75 from mRNA in a biological sample. The procedure involves sequencing at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement of position 556 as described in SEQ ID NO: 36, the position corresponding to or its complement of position 457 as described in SEQ ID NO: 41, the position corresponding to or its complement of position 378 as described in SEQ ID NO: 46, or the position corresponding to or its complement of position 617 as described in SEQ ID NO: 51. If the sequenced portion of the GPR75 nucleic acid molecule in the biological sample contains i) an adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3, ii) an adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, an adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, an adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, or an adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, or iii) an adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, an adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, an adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or an adenine at the position corresponding to position 617 as described in SEQ ID NO: 51, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0115] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement at position 5,911 as described in SEQ ID NO: 4; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement at position 910 as described in SEQ ID NO: 13, the position corresponding to or its complement at position 811 as described in SEQ ID NO: 18, the position corresponding to or its complement at position 732 as described in SEQ ID NO: 23, or the position corresponding to or its complement at position 971 as described in SEQ ID NO: 28; and / or iii) generating GPR75 from mRNA in a biological sample. The procedure involves sequencing at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement of position 910 as described in SEQ ID NO: 37, the position corresponding to or its complement of position 811 as described in SEQ ID NO: 42, the position corresponding to or its complement of position 732 as described in SEQ ID NO: 47, or the position corresponding to or its complement of position 971 as described in SEQ ID NO: 52. If the sequenced portion of the GPR75 nucleic acid molecule in the biological sample contains i) thymine at the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) uracil at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil at the position corresponding to position 971 as described in SEQ ID NO: 28, or iii) thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine at the position corresponding to position 971 as described in SEQ ID NO: 52, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0116] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes a position corresponding to or its complement to positions 5,920-5,923 as described in SEQ ID NO: 5; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes a position corresponding to or its complement to positions 919-922 as described in SEQ ID NO: 14, a position corresponding to or its complement to positions 820-823 as described in SEQ ID NO: 19, a position corresponding to or its complement to positions 741-744 as described in SEQ ID NO: 24, or a position corresponding to or its complement to positions 980-983 as described in SEQ ID NO: 29; and / or iii) generating GPR75 from mRNA in a biological sample. The procedure involves sequencing at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement to positions 919-922 as described in SEQ ID NO: 38, the position corresponding to or its complement to positions 820-823 as described in SEQ ID NO: 43, the position corresponding to or its complement to positions 741-744 as described in SEQ ID NO: 48, or the position corresponding to or its complement to positions 980-983 as described in SEQ ID NO: 53.The sequenced portion of the GPR75 nucleic acid molecule in the biological sample is either i) a deletion of AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1, ii) a deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, a deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, a deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or a deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10. If the GPR75 nucleic acid molecule in the biological sample contains a deletion of a tetranucleotide, or iii) a deletion of an AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, a deletion of an AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, a deletion of an AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or a deletion of an AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0117] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 6,411 as described in SEQ ID NO: 6; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 1,410 as described in SEQ ID NO: 15, the position corresponding to or its complement to position 1,311 as described in SEQ ID NO: 20, the position corresponding to or its complement to position 1,232 as described in SEQ ID NO: 25, or the position corresponding to or its complement to position 1,471 as described in SEQ ID NO: 30; and / or iii) generating GPR75 from mRNA in a biological sample. Sequence the nucleotide sequence of at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement of position 1,410 as described in SEQ ID NO: 39, the position corresponding to or its complement of position 1,311 as described in SEQ ID NO: 44, the position corresponding to or its complement of position 1,232 as described in SEQ ID NO: 49, or the position corresponding to or its complement of position 1,471 as described in SEQ ID NO: 54. If the sequenced portion of a GPR75 nucleic acid molecule in a biological sample contains i) a thymine insertion at the position corresponding to position 6,411 as described in SEQ ID NO: 6, ii) a uracil insertion at the position corresponding to position 1,410 as described in SEQ ID NO: 15, a uracil insertion at the position corresponding to position 1,311 as described in SEQ ID NO: 20, a uracil insertion at the position corresponding to position 1,232 as described in SEQ ID NO: 25, or a uracil insertion at the position corresponding to position 1,471 as described in SEQ ID NO: 30, or iii) a thymine insertion at the position corresponding to position 1,410 as described in SEQ ID NO: 39, a thymine insertion at the position corresponding to position 1,311 as described in SEQ ID NO: 44, a thymine insertion at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or a thymine insertion at the position corresponding to position 1,471 as described in SEQ ID NO: 54, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0118] In some embodiments, the determination step, detection step, or sequence analysis involves i) sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 5,831 as described in SEQ ID NO: 99; ii) sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement to position 830 as described in SEQ ID NO: 100, the position corresponding to or its complement to position 731 as described in SEQ ID NO: 101, the position corresponding to or its complement to position 652 as described in SEQ ID NO: 102, or the position corresponding to or its complement to position 891 as described in SEQ ID NO: 103; and / or iii) generating GPR75 from mRNA in a biological sample. The procedure involves sequencing at least a portion of the nucleotide sequence of a cDNA molecule, wherein the sequenced portion includes the position corresponding to or its complement at position 830 as described in SEQ ID NO: 104, the position corresponding to or its complement at position 731 as described in SEQ ID NO: 105, the position corresponding to or its complement at position 652 as described in SEQ ID NO: 106, or the position corresponding to or its complement at position 891 as described in SEQ ID NO: 107. If the sequenced portion of the GPR75 nucleic acid molecule in the biological sample contains i) guanine at the position corresponding to position 5,831 as described in SEQ ID NO: 99, ii) guanine at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine at the position corresponding to position 731 as described in SEQ ID NO: 101, guanine at the position corresponding to position 652 as described in SEQ ID NO: 102, or guanine at the position corresponding to position 891 as described in SEQ ID NO: 103, or iii) guanine at the position corresponding to position 830 as described in SEQ ID NO: 104, guanine at the position corresponding to position 731 as described in SEQ ID NO: 105, guanine at the position corresponding to position 652 as described in SEQ ID NO: 106, or guanine at the position corresponding to position 891 as described in SEQ ID NO: 107, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0119] In some embodiments, the determination step, detection step, or sequence analysis includes sequencing at least a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule in a biological sample, wherein the sequenced portion includes the position corresponding to or its complement at positions 5,540-5,546 as described in SEQ ID NO: 2, the position corresponding to or its complement at position 5,557 as described in SEQ ID NO: 3, the position corresponding to or its complement at position 5,911 as described in SEQ ID NO: 4, the position corresponding to or its complement at positions 5,920-5,923 as described in SEQ ID NO: 5, the position corresponding to or its complement at position 6,411 as described in SEQ ID NO: 6, or the position corresponding to or its complement at position 5,831 as described in SEQ ID NO: 99. If the sequenced portion of the GPR75 nucleic acid molecule in a biological sample contains a deletion of the CCAGTAG heptanucleotide at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, an adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3, a thymine at the position corresponding to position 5,911 as described in SEQ ID NO: 4, a deletion of the AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1, an insertion of thymine at the position corresponding to position 6,411 as described in SEQ ID NO: 6, or a guanine at the position corresponding to position 5,831 as described in SEQ ID NO: 99, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0120] In some embodiments, the determination step, detection step, or sequence analysis includes sequencing at least a portion of the nucleotide sequence of a GPR75 mRNA molecule in a biological sample, wherein the sequenced portion corresponds to the position or complement of positions 539-545 as described in SEQ ID NO: 11, the position or complement of positions 440-446 as described in SEQ ID NO: 16, the position or complement of positions 361-367 as described in SEQ ID NO: 21, the position or complement of positions 600-606 as described in SEQ ID NO: 26, the position or complement of position 556 as described in SEQ ID NO: 12, the position or complement of position 457 as described in SEQ ID NO: 17, the position or complement of position 378 as described in SEQ ID NO: 22, the position or complement of position 617 as described in SEQ ID NO: 27, the position or complement of position 910 as described in SEQ ID NO: 13, the position or complement of position 811 as described in SEQ ID NO: 18, the position or complement of position 732 as described in SEQ ID NO: 23, and 971 as described in SEQ ID NO: 28. This includes the position corresponding to the given position or its complement, the position corresponding to positions 919-922 as described in Sequence ID No. 14 or its complement, the position corresponding to positions 820-823 as described in Sequence ID No. 19 or its complement, the position corresponding to positions 741-744 as described in Sequence ID No. 24 or its complement, the position corresponding to positions 980-983 as described in Sequence ID No. 29 or its complement, the position corresponding to position 1,410 as described in Sequence ID No. 15 or its complement, the position corresponding to position 1,311 as described in Sequence ID No. 20 or its complement, the position corresponding to position 1,232 as described in Sequence ID No. 25 or its complement, the position corresponding to position 1,471 as described in Sequence ID No. 30 or its complement, the position corresponding to position 830 as described in Sequence ID No. 100 or its complement, the position corresponding to position 731 as described in Sequence ID No. 101 or its complement, the position corresponding to position 652 as described in Sequence ID No. 102 or its complement, and the position corresponding to position 891 as described in Sequence ID No. 103 or its complement.The sequenced portion of the GPR75 nucleic acid molecule in the biological sample lacks the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, or at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, or at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, or at the position corresponding to position 556 as described in SEQ ID NO: 12 It contains adenine at position 910, or adenine at position 457 as described in SEQ ID NO: 17, or adenine at position 378 as described in SEQ ID NO: 22, or adenine at position 617 as described in SEQ ID NO: 27, or uracil at position 910 as described in SEQ ID NO: 13, or uracil at position 811 as described in SEQ ID NO: 18, or uracil at position 732 as described in SEQ ID NO: 23, or uracil at position 971 as described in SEQ ID NO: 28, or uracil at positions 919-9 as described in SEQ ID NO: 7 It either lacks the AAAG tetranucleotide at the position corresponding to position 22, or lacks the AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, or lacks the AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or lacks the AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10, or includes an insertion of uracil at the position corresponding to position 1,410 as described in SEQ ID NO: 15, or includes an insertion of uracil at the position corresponding to position 1,311 as described in SEQ ID NO: 20, If a GPR75 nucleic acid molecule in a biological sample contains a uracil insertion at the position corresponding to position 1,232 as described in SEQ ID NO: 25, or a uracil insertion at the position corresponding to position 1,471 as described in SEQ ID NO: 30, or a guanine insertion at the position corresponding to position 830 as described in SEQ ID NO: 100, or a guanine insertion at the position corresponding to position 731 as described in SEQ ID NO: 101, or a guanine insertion at the position corresponding to position 652 as described in SEQ ID NO: 102, or a guanine insertion at the position corresponding to position 891 as described in SEQ ID NO: 103, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0121] In some embodiments, the determination step, detection step, or sequence analysis includes sequencing at least a portion of the nucleotide sequence of a GPR75 cDNA molecule generated from mRNA in a biological sample, wherein the sequenced portion corresponds to or is the position corresponding to positions 539-545 in SEQ ID NO: 35, positions corresponding to or is the position corresponding to positions 440-446 in SEQ ID NO: 40, positions corresponding to or is the position corresponding to positions 361-367 in SEQ ID NO: 45, positions corresponding to or is the position corresponding to positions 600-606 in SEQ ID NO: 50, positions corresponding to or is the position corresponding to position 556 in SEQ ID NO: 36, positions corresponding to or is the position corresponding to position 457 in SEQ ID NO: 41, positions corresponding to or is the position corresponding to position 378 in SEQ ID NO: 46, positions corresponding to or is the position corresponding to position 617 in SEQ ID NO: 51, positions corresponding to or is the position corresponding to position 910 in SEQ ID NO: 37, positions corresponding to or is the position corresponding to position 811 in SEQ ID NO: 42, positions corresponding to or is the position corresponding to position 732 in SEQ ID NO: 47, and positions corresponding to position 971 in SEQ ID NO: 52 This includes the position corresponding to or its complement, the position corresponding to positions 919-922 in Sequence ID No. 38 or its complement, the position corresponding to positions 820-823 in Sequence ID No. 43 or its complement, the position corresponding to positions 741-744 in Sequence ID No. 48 or its complement, the position corresponding to positions 980-983 in Sequence ID No. 53 or its complement, the position corresponding to position 1,410 in Sequence ID No. 39 or its complement, the position corresponding to position 1,311 in Sequence ID No. 44 or its complement, the position corresponding to position 1,232 in Sequence ID No. 49 or its complement, the position corresponding to position 1,471 in Sequence ID No. 54, the position corresponding to position 830 in Sequence ID No. 104 or its complement, the position corresponding to position 731 in Sequence ID No. 105 or its complement, the position corresponding to position 652 in Sequence ID No. 106 or its complement, or the position corresponding to position 891 in Sequence ID No. 107.The sequenced portion of the GPR75 nucleic acid molecule in the biological sample lacks a CCAGTAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, or at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, or at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, or at position 556 as described in SEQ ID NO: 36 It contains adenine at the corresponding position, or adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, or adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 51, or thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, or thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, or thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine at the position corresponding to position 971 as described in SEQ ID NO: 52, or thymine at position 919~ as described in SEQ ID NO: 31 Either the AAAG tetranucleotide at position 922 is missing, or the AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 32 is missing, or the AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 33 is missing, or the AAAG tetranucleotide at positions 980-983 as described in SEQ ID NO: 34 is missing, or the thymine insertion at position 1,410 as described in SEQ ID NO: 39 is included, or the thymine insertion at position 1,311 as described in SEQ ID NO: 44 is included. If the GPR75 nucleic acid molecule in the biological sample contains a thymine insertion at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or a thymine insertion at the position corresponding to position 1,471 as described in SEQ ID NO: 54, or a guanine insertion at the position corresponding to position 830 as described in SEQ ID NO: 104, or a guanine insertion at the position corresponding to position 731 as described in SEQ ID NO: 105, or a guanine insertion at the position corresponding to position 652 as described in SEQ ID NO: 106, or a guanine insertion at the position corresponding to position 891 as described in SEQ ID NO: 107, then the GPR75 nucleic acid molecule in the biological sample is a GPR75 missense variant nucleic acid molecule.

[0122] In some embodiments, the determination step, detection step, or sequence analysis involves a) i) a portion of the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to the position corresponding to positions 5,540-5,546 described in SEQ ID NO: 2, ii) a portion of the nucleotide sequence of the GPR75 mRNA molecule adjacent to the position corresponding to positions 539-545 described in SEQ ID NO: 11, positions 440-446 described in SEQ ID NO: 16, positions 361-367 described in SEQ ID NO: 21, or positions 600-606 described in SEQ ID NO: 26, and / or iii) GPR75 adjacent to the position corresponding to positions 539-545 described in SEQ ID NO: 35, positions 440-446 described in SEQ ID NO: 40, positions 361-367 described in SEQ ID NO: 45, or positions 600-606 described in SEQ ID NO: 50. The process involves: (a) contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of a cDNA molecule; (b) extending the primer via at least the following locations: i) the nucleotide sequence of the GPR75 genome nucleic acid molecule corresponding to positions 5,540-5,546 described in SEQ ID NO: 2, ii) the nucleotide sequence of the GPR75 mRNA molecule corresponding to positions 539-545 described in SEQ ID NO: 11, positions 440-446 described in SEQ ID NO: 16, positions 361-367 described in SEQ ID NO: 21, or positions 600-606 described in SEQ ID NO: 26, and / or iii) the nucleotide sequence of the GPR75 cDNA molecule corresponding to positions 539-545 described in SEQ ID NO: 35, positions 440-446 described in SEQ ID NO: 40, positions 361-367 described in SEQ ID NO: 45, or positions 600-606 described in SEQ ID NO: 50; and (c) the extension product of the primer is i) 5,540-5,ii) Deletion of the CCAGUAG heptanucleotide at the position corresponding to position 546, ii) Deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, Deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, Deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or Deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10 This includes determining whether the following are included: iii) deletion of CCAGTAG heptanucleotide at positions corresponding to positions 539-545 as described in SEQ ID NO: 31, deletion of CCAGTAG heptanucleotide at positions corresponding to positions 440-446 as described in SEQ ID NO: 32, deletion of CCAGTAG heptanucleotide at positions corresponding to positions 361-367 as described in SEQ ID NO: 33, or deletion of CCAGTAG heptanucleotide at positions corresponding to positions 600-606 as described in SEQ ID NO: 34.

[0123] In some embodiments, the determination step, detection step, or sequence analysis involves contacting a biological sample with a primer that hybridizes with a) i) a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule adjacent to the position corresponding to position 5,557 as described in SEQ ID NO: 3, ii) a portion of the nucleotide sequence of a GPR75 mRNA molecule adjacent to the position corresponding to position 556 as described in SEQ ID NO: 12, position 457 as described in SEQ ID NO: 17, position 378 as described in SEQ ID NO: 22, position 617 as described in SEQ ID NO: 27, and / or iii) a portion of the nucleotide sequence of a GPR75 cDNA molecule adjacent to the position corresponding to position 556 as described in SEQ ID NO: 36, position 457 as described in SEQ ID NO: 41, position 378 as described in SEQ ID NO: 46, or position 617 as described in SEQ ID NO: 51, and b) i) the position of the nucleotide sequence of a GPR75 genomic nucleic acid molecule corresponding to position 5,557 as described in SEQ ID NO: 3, ii) the GPR75 corresponding to position 556 as described in SEQ ID NO: 12, position 457 as described in SEQ ID NO: 17, position 378 as described in SEQ ID NO: 22, or position 617 as described in SEQ ID NO: 27 The method includes: extending the primer via at least the position of the nucleotide sequence of the mRNA molecule and / or iii) the position of the nucleotide sequence of the GPR75 cDNA molecule corresponding to position 556 as described in SEQ ID NO: 36, position 457 as described in SEQ ID NO: 41, position 378 as described in SEQ ID NO: 46, or position 617 as described in SEQ ID NO: 51; and c) determining whether the primer extension product contains i) adenine corresponding to position 5,557 as described in SEQ ID NO: 3, ii) adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, or iii) adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 51.

[0124] In some embodiments, the determination step, detection step, or sequence analysis involves contacting a biological sample with a primer that hybridizes with a) i) a portion of the nucleotide sequence of a GPR75 genomic nucleic acid molecule adjacent to the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) a portion of the nucleotide sequence of a GPR75 mRNA molecule adjacent to the position corresponding to position 910 as described in SEQ ID NO: 13, position 811 as described in SEQ ID NO: 18, position 732 as described in SEQ ID NO: 23, or position 971 as described in SEQ ID NO: 28, and / or iii) a portion of the nucleotide sequence of a GPR75 cDNA molecule adjacent to the position corresponding to position 910 as described in SEQ ID NO: 37, position 811 as described in SEQ ID NO: 42, position 732 as described in SEQ ID NO: 47, or position 971 as described in SEQ ID NO: 52, and b) i) the position of the nucleotide sequence of a GPR75 genomic nucleic acid molecule corresponding to position 5,911 as described in SEQ ID NO: 4, and ii) the GPR75 corresponding to position 910 as described in SEQ ID NO: 13, position 811 as described in SEQ ID NO: 18, position 732 as described in SEQ ID NO: 23, or position 971 as described in SEQ ID NO: 28 The method includes: extending the primer via at least the position of the nucleotide sequence of the mRNA molecule and / or iii) the position of the nucleotide sequence of the GPR75 cDNA molecule corresponding to position 910 as described in SEQ ID NO: 37, position 811 as described in SEQ ID NO: 42, position 732 as described in SEQ ID NO: 47, or position 971 as described in SEQ ID NO: 52; and c) determining whether the primer extension product contains i) thymine at the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) uracil at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil at the position corresponding to position 971 as described in SEQ ID NO: 28, or iii) thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine at the position corresponding to position 971 as described in SEQ ID NO: 52.

[0125] In some embodiments, the determination step, detection step, or sequence analysis involves a) i) a portion of the nucleotide sequence of the GPR75 genome nucleic acid molecule adjacent to the position corresponding to positions 5,920-5,923 described in SEQ ID NO: 5, ii) a portion of the nucleotide sequence of the GPR75 mRNA molecule adjacent to the position corresponding to positions 919-922 described in SEQ ID NO: 14, positions 820-823 described in SEQ ID NO: 19, positions 741-744 described in SEQ ID NO: 24, or positions 980-983 described in SEQ ID NO: 29, and / or iii) GPR75 adjacent to the position corresponding to positions 919-922 described in SEQ ID NO: 38, positions 820-823 described in SEQ ID NO: 43, positions 741-744 described in SEQ ID NO: 48, or positions 980-983 described in SEQ ID NO: 53. The process involves: a) contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of a cDNA molecule; and extending the primer via at least the following locations: i) the nucleotide sequence of the GPR75 genome nucleic acid molecule corresponding to positions 5,920-5,923 described in SEQ ID NO: 5, ii) the nucleotide sequence of the GPR75 mRNA molecule corresponding to positions 919-922 described in SEQ ID NO: 14, positions 820-823 described in SEQ ID NO: 19, positions 741-744 described in SEQ ID NO: 24, or positions 980-983 described in SEQ ID NO: 29, and / or iii) the nucleotide sequence of the GPR75 cDNA molecule corresponding to positions 919-922 described in SEQ ID NO: 38, positions 820-823 described in SEQ ID NO: 43, positions 741-744 described in SEQ ID NO: 48, or positions 980-983 described in SEQ ID NO: 53; and c) the extension product of the primer is i) 5,920-5,This includes determining whether the following are included: ii) a deletion of AAAG tetranucleotide at the position corresponding to position 923, ii) a deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, a deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, a deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or a deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10, or iii) a deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, a deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, a deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or a deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 34.

[0126] In some embodiments, the determination step, detection step, or sequence analysis involves a) i) a portion of the nucleotide sequence of the GPR75 genome nucleic acid molecule adjacent to the position corresponding to position 6,411 as described in SEQ ID NO: 6, ii) a portion of the nucleotide sequence of the GPR75 mRNA molecule adjacent to the position corresponding to position 1,410 as described in SEQ ID NO: 15, position 1,311 as described in SEQ ID NO: 20, position 1,232 as described in SEQ ID NO: 25, or position 1,471 as described in SEQ ID NO: 30, and / or iii) GPR75 adjacent to the position corresponding to position 1,410 as described in SEQ ID NO: 39, position 1,311 as described in SEQ ID NO: 44, position 1,232 as described in SEQ ID NO: 49, or position 1,471 as described in SEQ ID NO: 54. The process involves contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of a cDNA molecule, and b)i) the position of the nucleotide sequence of the GPR75 genome nucleic acid molecule corresponding to position 6,411 as described in SEQ ID NO: 6, ii) the position of the nucleotide sequence of the GPR75 mRNA molecule corresponding to position 1,410 as described in SEQ ID NO: 15, position 1,311 as described in SEQ ID NO: 20, position 1,232 as described in SEQ ID NO: 25, or position 1,471 as described in SEQ ID NO: 30, and / or iii) the GPR75 corresponding to position 1,410 as described in SEQ ID NO: 39, position 1,311 as described in SEQ ID NO: 44, position 1,232 as described in SEQ ID NO: 49, or position 1,471 as described in SEQ ID NO: 54. The method includes: a) extending the primer at least via the nucleotide sequence of a cDNA molecule; and determining whether the primer extension product includes i) an insertion of thymine at the position corresponding to position 6,411 as described in SEQ ID NO: 6; ii) an insertion of uracil at the position corresponding to position 1,410 as described in SEQ ID NO: 15; an insertion of uracil at the position corresponding to position 1,311 as described in SEQ ID NO: 20; an insertion of uracil at the position corresponding to position 1,232 as described in SEQ ID NO: 25; or an insertion of uracil at the position corresponding to position 1,471 as described in SEQ ID NO: 30; or iii) an insertion of thymine at the position corresponding to position 1,410 as described in SEQ ID NO: 39; an insertion of thymine at the position corresponding to position 1,311 as described in SEQ ID NO: 44; an insertion of thymine at the position corresponding to position 1,232 as described in SEQ ID NO: 49; or an insertion of thymine at the position corresponding to position 1,471 as described in SEQ ID NO: 54.

[0127] In some embodiments, the determination step, detection step, or sequence analysis involves a) i) a portion of the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to the position corresponding to position 5,831 as described in SEQ ID NO: 99, ii) a portion of the nucleotide sequence of the GPR75 mRNA molecule adjacent to the position corresponding to position 830 as described in SEQ ID NO: 100, position 731 as described in SEQ ID NO: 101, position 652 as described in SEQ ID NO: 102, or position 891 as described in SEQ ID NO: 103, and / or iii) GPR75 adjacent to the position corresponding to position 830 as described in SEQ ID NO: 104, position 731 as described in SEQ ID NO: 105, position 652 as described in SEQ ID NO: 106, or position 891 as described in SEQ ID NO: 107 The process involves: (i) bringing a biological sample into contact with a primer that hybridizes with a portion of the nucleotide sequence of a cDNA molecule; (ii) extending the primer via at least the following locations: i) the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to the position corresponding to position 5,831 described in SEQ ID NO: 99; ii) the nucleotide sequence of the GPR75 mRNA molecule adjacent to the position corresponding to position 830 described in SEQ ID NO: 100, position 731 described in SEQ ID NO: 101, position 652 described in SEQ ID NO: 102, or position 891 described in SEQ ID NO: 103; and (iii) extending the primer via at least the following locations: i) the nucleotide sequence of the GPR75 cDNA molecule adjacent to the position corresponding to position 830 described in SEQ ID NO: 104, position 731 described in SEQ ID NO: 105, position 652 described in SEQ ID NO: 106, or position 891 described in SEQ ID NO: 107; and (iii) the extension product of the primer is i) the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to position 5,831 described in SEQ ID NO: 99.This includes determining whether the following are included: ii) guanine or its complement at the position corresponding to 831, ii) guanine or its complement at the position corresponding to 830 as described in SEQ ID NO: 100, guanine or its complement at the position corresponding to 731 as described in SEQ ID NO: 101, guanine or its complement at the position corresponding to 652 as described in SEQ ID NO: 102, or guanine or its complement at the position corresponding to 891 as described in SEQ ID NO: 103, or iii) guanine or its complement at the position corresponding to 830 as described in SEQ ID NO: 104, guanine or its complement at the position corresponding to 731 as described in SEQ ID NO: 105, guanine or its complement at the position corresponding to 652 as described in SEQ ID NO: 106, or guanine or its complement at the position corresponding to 891 as described in SEQ ID NO: 107.

[0128] In some embodiments, the determination step, detection step, or sequence analysis involves a) contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to the positions corresponding to positions 5,540-5,546 in SEQ ID NO: 2, position 5,557 in SEQ ID NO: 3, position 5,911 in SEQ ID NO: 4, positions 5,920-5,923 in SEQ ID NO: 5, position 6,411 in SEQ ID NO: 6, or position 5,831 in SEQ ID NO: 99, and b) contacting a primer that hybridizes with a portion of the nucleotide sequence of the GPR75 genomic nucleic acid molecule adjacent to the positions corresponding to positions 5,540-5,546 in SEQ ID NO: 2, position 5,557 in SEQ ID NO: 3, position 5,911 in SEQ ID NO: 4, positions 5,920-5,923 in SEQ ID NO: 5, position 6,411 in SEQ ID NO: 6, or position 5,831 in SEQ ID NO: 99 The method includes: a) extending the primer at least via the nucleotide sequence of the GPR75 genome nucleic acid molecule; and determining whether the extended primer product lacks the CCAGTAG heptanucleotide at the position corresponding to positions 5,540-5,546 described in SEQ ID NO: 1, contains adenine at the position corresponding to position 5,557 described in SEQ ID NO: 3, contains thymine at the position corresponding to position 5,911 described in SEQ ID NO: 4, lacks the AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 described in SEQ ID NO: 1, contains a thymine insertion at the position corresponding to position 6,411 described in SEQ ID NO: 6, or contains guanine or its complement at the position corresponding to position 5,831 described in SEQ ID NO: 99.

[0129] In some embodiments, the determination step, detection step, or sequence analysis is performed by a) positions 539-545 in SEQ ID NO: 11, positions 440-446 in SEQ ID NO: 16, positions 361-367 in SEQ ID NO: 21, positions 600-606 in SEQ ID NO: 26, position 556 in SEQ ID NO: 12, position 457 in SEQ ID NO: 17, position 378 in SEQ ID NO: 22, position 617 in SEQ ID NO: 27, position 910 in SEQ ID NO: 13, position 811 in SEQ ID NO: 18, position 732 in SEQ ID NO: 23, position 97 in SEQ ID NO: 28 GPR75 adjacent to the position corresponding to position 1, positions 919-922 in SEQ ID NO: 14, positions 820-823 in SEQ ID NO: 19, positions 741-744 in SEQ ID NO: 24, positions 980-983 in SEQ ID NO: 29, position 1,410 in SEQ ID NO: 15, position 1,311 in SEQ ID NO: 20, position 1,232 in SEQ ID NO: 25, position 1,471 in SEQ ID NO: 30, position 830 in SEQ ID NO: 100, position 731 in SEQ ID NO: 101, position 652 in SEQ ID NO: 102, or position 891 in SEQ ID NO: 103 b) Contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of an mRNA molecule, and position 539-545 in SEQ ID NO: 11, position 440-446 in SEQ ID NO: 16, position 361-367 in SEQ ID NO: 21, position 600-606 in SEQ ID NO: 26, position 556 in SEQ ID NO: 12, position 457 in SEQ ID NO: 17, position 378 in SEQ ID NO: 22, position 617 in SEQ ID NO: 27, position 910 in SEQ ID NO: 13, position 811 in SEQ ID NO: 18, position 732 in SEQ ID NO: 23, position 971 in SEQ ID NO: 28, position 919-922 in SEQ ID NO: 14, position 820-823 in SEQ ID NO: 19, position 741-744 in SEQ ID NO: 24, position 980-983 in SEQ ID NO: 29, position 1,410 in SEQ ID NO: 15, position 1,311 in SEQ ID NO: 20, position 1,232 in SEQ ID NO: 25, position 1,GPR75 corresponding to position 471, position 830 in sequence number 100, position 731 in sequence number 101, position 652 in sequence number 102, or position 891 in sequence number 103. c) extending the primer at least via the nucleotide sequence of the mRNA molecule, and c) whether the extended primer product lacks the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, whether it lacks the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, whether it lacks the CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, whether it lacks the CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, whether it contains adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, whether it contains adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, whether it contains adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, whether it contains adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, and whether it contains uracil at the position corresponding to position 910 as described in SEQ ID NO: 13. Whether or not, whether or not it contains uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, whether or not it contains uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, whether or not it contains uracil at the position corresponding to position 971 as described in SEQ ID NO: 28, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 820-823 as described in SEQ ID NO: 8, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 741-744 as described in SEQ ID NO: 9, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 980-983 as described in SEQ ID NO: 10, whether or not it includes an insertion of uracil at the position corresponding to position 1,410 as described in SEQ ID NO: 15, whether or not it includes an insertion of uracil at the position corresponding to position 1,311 as described in SEQ ID NO: 20, whether or not it includes an insertion of uracil at the position corresponding to position 1,232 as described in SEQ ID NO: 25, 1,This includes determining whether or not it includes an insertion of uracil at position 471, whether or not it includes guanine at position 830 as described in SEQ ID NO: 100, whether or not it includes guanine at position 731 as described in SEQ ID NO: 101, whether or not it includes guanine at position 652 as described in SEQ ID NO: 102, or whether or not it includes guanine at position 891 as described in SEQ ID NO: 103.

[0130] In some embodiments, the determination step, detection step, or sequence analysis is performed by a) positions 539-545 in SEQ ID NO: 35, positions 440-446 in SEQ ID NO: 40, positions 361-367 in SEQ ID NO: 45, positions 600-606 in SEQ ID NO: 50, position 556 in SEQ ID NO: 36, position 457 in SEQ ID NO: 41, position 378 in SEQ ID NO: 46, position 617 in SEQ ID NO: 51, position 910 in SEQ ID NO: 37, position 811 in SEQ ID NO: 42, position 732 in SEQ ID NO: 47, position 97 in SEQ ID NO: 52 GPR75 adjacent to the position corresponding to position 1, positions 919-922 in SEQ ID NO: 38, positions 820-823 in SEQ ID NO: 43, positions 741-744 in SEQ ID NO: 48, positions 980-983 in SEQ ID NO: 53, position 1,410 in SEQ ID NO: 39, position 1,311 in SEQ ID NO: 44, position 1,232 in SEQ ID NO: 49, position 1,471 in SEQ ID NO: 54, position 830 in SEQ ID NO: 104, position 731 in SEQ ID NO: 105, position 652 in SEQ ID NO: 106, or position 891 in SEQ ID NO: 107 b) Contacting a biological sample with a primer that hybridizes with a portion of the nucleotide sequence of a cDNA molecule, and position 539-545 as described in SEQ ID NO: 35, position 440-446 as described in SEQ ID NO: 40, position 361-367 as described in SEQ ID NO: 45, position 600-606 as described in SEQ ID NO: 50, position 556 as described in SEQ ID NO: 36, position 457 as described in SEQ ID NO: 41, position 378 as described in SEQ ID NO: 46, position 617 as described in SEQ ID NO: 51, position 910 as described in SEQ ID NO: 37, position 811 as described in SEQ ID NO: 42, position 732 as described in SEQ ID NO: 47, position 971 as described in SEQ ID NO: 52, position 919-922 as described in SEQ ID NO: 38, position 820-823 as described in SEQ ID NO: 43, position 741-744 as described in SEQ ID NO: 48, position 980-983 as described in SEQ ID NO: 53, position 1,410 as described in SEQ ID NO: 39, position 1,311 as described in SEQ ID NO: 44, position 1,232 as described in SEQ ID NO: 49, position 1,GPR75 corresponding to position 471, position 830 in sequence number 104, position 731 in sequence number 105, position 652 in sequence number 106, or position 891 in sequence number 107. c) extending the primer at least via the nucleotide sequence of the cDNA molecule, and c) whether the extended primer product lacks the CCAGTAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, whether it lacks the CCAGTAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, whether it lacks the CCAGTAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, whether it lacks the CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, whether it contains adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, whether it contains adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, whether it contains adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, whether it contains adenine at the position corresponding to position 617 as described in SEQ ID NO: 51, and whether it contains thymine at the position corresponding to position 910 as described in SEQ ID NO: 37 Whether or not, whether or not it contains thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, whether or not it contains thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, whether or not it contains thymine at the position corresponding to position 971 as described in SEQ ID NO: 52, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 741-744 as described in SEQ ID NO: 33, whether or not it lacks AAAG tetranucleotides at the positions corresponding to positions 980-983 as described in SEQ ID NO: 34, whether or not it includes an insertion of thymine at the position corresponding to position 1,410 as described in SEQ ID NO: 39, whether or not it includes an insertion of thymine at the position corresponding to position 1,311 as described in SEQ ID NO: 44, whether or not it includes an insertion of thymine at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or the 1,This includes determining whether or not it includes an insertion of thymine at position 471, whether or not it includes guanine at position 830 as described in SEQ ID NO: 104, whether or not it includes guanine at position 731 as described in SEQ ID NO: 105, whether or not it includes guanine at position 652 as described in SEQ ID NO: 106, or whether or not it includes guanine at position 891 as described in SEQ ID NO: 107.

[0131] In some embodiments, the assay includes sequencing the entire nucleic acid molecule. In some embodiments, only the GPR75 genomic nucleic acid molecule is analyzed. In some embodiments, only the GPR75 mRNA is analyzed. In some embodiments, only the GPR75 cDNA obtained from the GPR75 mRNA is analyzed.

[0132] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding the human GPR75 polypeptide, wherein the amplified portion is i) a deletion of the CCAGUAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, ii) a deletion of the CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, a deletion of the CCAGUAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, a deletion of the CCAGUAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10 a) Amplification including the deletion of ocide or its complement, and / or iii) deletion of CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, deletion of CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, deletion of CCAGTAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or deletion of CCAGTAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, b) labeling the amplified nucleic acid molecule with a detectable label, and c) contacting the labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe is i) 5,540-5, as described in SEQ ID NO: 1ii) Lack of CCAGUAG heptanucleotide or its complement at the position corresponding to position 546, ii) Lack of CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, Lack of CCAGUAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, Lack of CCAGUAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or Lack of CCAGUAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, and / or iii) 53 as described in SEQ ID NO: 31 d) Contacting a nucleotide sequence that hybridizes under stringent conditions with an amplified nucleic acid molecule lacking a CCAGTAG heptanucleotide or its complement at positions 9-545, lacking a CCAGTAG heptanucleotide or its complement at positions 440-446 as described in SEQ ID NO: 32, lacking a CCAGTAG heptanucleotide or its complement at positions 361-367 as described in SEQ ID NO: 33, or lacking a CCAGTAG heptanucleotide or its complement at positions 600-606 as described in SEQ ID NO: 34; and d) detecting a detectable label.

[0133] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion is i) adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, ii) adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, or adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22. a) adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, and / or iii) adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, a) amplifying the amplified nucleic acid molecule with a detectable label, and c) labeling The nucleic acid molecule is brought into contact with a support containing a change-specific probe, wherein the change-specific probe is i) adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, ii) adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22, or adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, and iii) Contacting a nucleic acid sequence of an amplified nucleic acid molecule containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, with a nucleotide sequence that hybridizes under stringent conditions; and d) detecting a detectable label.

[0134] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion includes i) thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, and / or iii) thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, and b) labeling the amplified nucleic acid molecule with a detectable label, and c) The method involves contacting a labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe is i) thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28. , and / or iii) contacting a nucleotide sequence that hybridizes under stringent conditions with a nucleic acid sequence of an amplified nucleic acid molecule containing thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52; and d) detecting a detectable label.

[0135] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion is i) a deletion of an AAAG tetranucleotide or its complement at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1, ii) a deletion of an AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, a deletion of an AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, a deletion of an AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or an AAAG tetranucleo at the position corresponding to positions 980-983 as described in SEQ ID NO: 10 a) Amplification including deletion of tide or its complement, and / or iii) deletion of AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, deletion of AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, deletion of AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or deletion of AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, b) labeling the amplified nucleic acid molecule with a detectable label, and c) contacting the labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe is i) 5,920-5, as described in SEQ ID NO: 1ii) Lacking an AAAG tetranucleotide or its complement at the position corresponding to position 923, as described in SEQ ID NO: 7, as well as a lack of an AAAG tetranucleotide or its complement at the position corresponding to positions 919-922, as described in SEQ ID NO: 8, as well as a lack of an AAAG tetranucleotide or its complement at the position corresponding to positions 820-823, as described in SEQ ID NO: 9, as well as a lack of an AAAG tetranucleotide or its complement at the position corresponding to positions 741-744, as described in SEQ ID NO: 9, or as described in SEQ ID NO: 10, as well as a lack of an AAAG tetranucleotide or its complement at the position corresponding to positions 980-983, and / or iii) 919 as described in SEQ ID NO: 31 d) Contacting a nucleotide sequence that hybridizes under stringent conditions with an amplified nucleic acid molecule lacking an AAAG tetranucleotide or its complement at the position corresponding to ~922, lacking an AAAG tetranucleotide or its complement at the position corresponding to 820~823 as described in SEQ ID NO: 32, lacking an AAAG tetranucleotide or its complement at the position corresponding to 741~744 as described in SEQ ID NO: 33, or lacking an AAAG tetranucleotide or its complement at the position corresponding to 980~983 as described in SEQ ID NO: 34; and d) detecting a detectable label.

[0136] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion includes i) an insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, ii) an insertion of uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15, an insertion of uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20, an insertion of uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, an insertion of uracil or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 30, and / or iii) an insertion of thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, an insertion of thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, an insertion of thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or thymine at the position corresponding to position 1,471 as described in SEQ ID NO: 54 a) amplification, including insertion of its complement, b) labeling the amplified nucleic acid molecule with a detectable label, and c) contacting the labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe is i) insertion of thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, ii) insertion of uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15, or insertion of uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20, sequence Insertion of uracil or its complement at the position corresponding to position 1,232 as described in number 25, or insertion of uracil or its complement at the position corresponding to position 1,471 as described in sequence number 30, and / or iii) Insertion of thymine or its complement at the position corresponding to position 1,410 as described in sequence number 39, insertion of thymine or its complement at the position corresponding to position 1,311 as described in sequence number 44, insertion of thymine or its complement at the position corresponding to position 1,232 as described in sequence number 49, or 1, as described in sequence number 54d) contacting a nucleic acid sequence of an amplified nucleic acid molecule containing an insertion of thymine or its complement at the position corresponding to 471 with a nucleotide sequence that hybridizes under stringent conditions, and d) detecting a detectable label.

[0137] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion is i) guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99, ii) guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, or guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102. a) Amplifying the guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103, and / or iii) amplifying the guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, the guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, the guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or the guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107, a) labeling the amplified nucleic acid molecule with a detectable label, and c) The method involves contacting a labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe is i) guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99, ii) guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103. and / or iii) contacting a nucleic acid sequence of an amplified nucleic acid molecule containing guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107, with a nucleotide sequence that hybridizes under stringent conditions, and d) detecting a detectable label.

[0138] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion lacks a CCAGTAG heptanucleotide or its complement at the position corresponding to 5,540-5,546 as described in SEQ ID NO: 1, contains adenine or its complement at the position corresponding to 5,557 as described in SEQ ID NO: 3, contains thymine or its complement at the position corresponding to 5,911 as described in SEQ ID NO: 4, lacks an AAAG tetranucleotide or its complement at the position corresponding to 5,920-5,923 as described in SEQ ID NO: 1, contains an insertion of thymine or its complement at the position corresponding to 6,411 as described in SEQ ID NO: 6, or contains guanine or its complement at the position corresponding to 5,831 as described in SEQ ID NO: 99, and b) labeling the amplified nucleic acid molecule with a detectable label, and c) d) Contacting a labeled nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe includes a nucleotide sequence that hybridizes under stringent conditions with the nucleic acid sequence of an amplified nucleic acid molecule containing a CCAGTAG heptanucleotide or its complement at positions 5,540-5,546 as described in SEQ ID NO: 1, adenine or its complement at position 5,557 as described in SEQ ID NO: 3, thymine or its complement at position 5,911 as described in SEQ ID NO: 4, AAAG tetranucleotide or its complement at positions 5,920-5,923 as described in SEQ ID NO: 1, an insertion of thymine or its complement at position 6,411 as described in SEQ ID NO: 6, or guanine or its complement at position 5,831 as described in SEQ ID NO: 99; and d) detecting a detectable label.

[0139] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion lacks the CCAGUAG heptanucleotide or its complement at the positions corresponding to positions 539-545 as described in SEQ ID NO: 7, the CCAGUAG heptanucleotide or its complement at the positions corresponding to positions 440-446 as described in SEQ ID NO: 8, or the CCAGUAG heptanucleotide at the positions corresponding to positions 361-367 as described in SEQ ID NO: 9. Or lacking its complement, the CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, or lacking its complement, containing adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, containing adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, containing adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, containing adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, or at position 910 as described in SEQ ID NO: 13 Contains uracil or its complement at the corresponding position, contains uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, contains uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, contains uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 8 It includes: lacking AAAG tetranucleotide or its complement at positions 741-744 as described in SEQ ID NO: 9; lacking AAAG tetranucleotide or its complement at positions 980-983 as described in SEQ ID NO: 10; including an insertion of uracil or its complement at position 1,410 as described in SEQ ID NO: 15; including an insertion of uracil or its complement at position 1,311 as described in SEQ ID NO: 20; including an insertion of uracil or its complement at position 1,232 as described in SEQ ID NO: 25.a) Amplifying the nucleic acid molecule, including the insertion of uracil or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 30, including guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, including guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, including guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or including guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 103; b) labeling the amplified nucleic acid molecule with a detectable label; c) labeling The method involves contacting a nucleic acid molecule with a support containing a change-specific probe, wherein the change-specific probe lacks CCAGUAG heptanucleotide or its complement at positions corresponding to 539-545 as described in SEQ ID NO: 7, CCAGUAG heptanucleotide or its complement at positions corresponding to 440-446 as described in SEQ ID NO: 8, CCAGUAG heptanucleotide or its complement at positions corresponding to 361-367 as described in SEQ ID NO: 9, and CCAGUAG heptanucleotide at positions corresponding to 600-606 as described in SEQ ID NO: 10. Lacking a ptanucleotide or its complement, containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, containing adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, containing adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22, containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 27, containing uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, containing uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18 Uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, which includes its complement, or uracil at the position corresponding to position 971 as described in SEQ ID NO: 28, which includes its complement, or lacking AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, lacking AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, lacking AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 9,This includes contacting a nucleotide sequence that hybridizes under stringent conditions with an amplified nucleic acid molecule, which lacks the AAAG tetranucleotide or its complement at positions 980-983 as described in SEQ ID NO: 10, includes an insertion of uracil or its complement at position 1,410 as described in SEQ ID NO: 15, includes an insertion of uracil or its complement at position 1,311 as described in SEQ ID NO: 20, includes an insertion of uracil or its complement at position 1,232 as described in SEQ ID NO: 25, includes an insertion of uracil or its complement at position 1,471 as described in SEQ ID NO: 30, includes guanine or its complement at position 830 as described in SEQ ID NO: 100, includes guanine or its complement at position 731 as described in SEQ ID NO: 101, includes guanine or its complement at position 652 as described in SEQ ID NO: 102, or includes guanine or its complement at position 891 as described in SEQ ID NO: 103.

[0140] In some embodiments, the determination step, detection step, or sequence analysis involves a) amplifying at least a portion of a nucleic acid molecule encoding a human GPR75 polypeptide, wherein the amplified portion lacks a CCAGTAG heptanucleotide or its complement at positions corresponding to positions 539-545 as described in SEQ ID NO: 31, a CCAGTAG heptanucleotide or its complement at positions corresponding to positions 440-446 as described in SEQ ID NO: 32, or a CCAGTAG heptanucleotide at positions corresponding to positions 361-367 as described in SEQ ID NO: 33. Lack of rheotide or its complement, lacking the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, containing adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, containing adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, and the 9 as described in SEQ ID NO: 37 The AAAG tetranucleotide or its complement at the position corresponding to position 820-823 as described in SEQ ID NO: 32, which includes thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, which includes thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, which includes thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, which lacks the AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31 Lacking, lacking AAAG tetranucleotide or its complement at positions 741-744 as described in SEQ ID NO: 33, lacking AAAG tetranucleotide or its complement at positions 980-983 as described in SEQ ID NO: 34, including insertion of thymine or its complement at position 1,410 as described in SEQ ID NO: 39, including insertion of thymine or its complement at position 1,311 as described in SEQ ID NO: 44, including insertion of thymine or its complement at position 1,232 as described in SEQ ID NO: 49,a) Amplification including the insertion of thymine or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 54, including guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, including guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, including guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or including guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107; b) labeling the amplified nucleic acid molecule with a detectable label; c) labeling the nucleic acid The molecule is brought into contact with a support containing a change-specific probe, wherein the change-specific probe lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, and the CCAGTA at the position corresponding to positions 600-606 as described in SEQ ID NO: 34. Lacking G heptanucleotide or its complement, containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, containing adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, containing adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, containing thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, containing thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42 The thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, which includes its complement, or the thymine at the position corresponding to position 971 as described in SEQ ID NO: 52, which includes its complement, or the AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, which lacks the AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, which lacks the AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 33,The process includes contacting a nucleic acid sequence of an amplified nucleic acid molecule containing a nucleotide sequence that hybridizes under stringent conditions, and d) detecting a detectable label.

[0141] In some embodiments, the nucleic acid molecule is mRNA, and the determination step further includes reverse transcribing the mRNA into cDNA before the amplification step. In some embodiments, the determination step, detection step, or sequence analysis involves contacting nucleic acid molecules in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe is i) lacking a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, ii) lacking a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, lacking a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, lacking a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, or lacking a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in SEQ ID NO: 10 The method comprises contacting a nucleotide sequence of an amplified nucleic acid molecule that lacks a CCAGUAG heptanucleotide or its complement, and / or iii) lacks a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, lacks a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, lacks a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or lacks a CCAGUAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0142] In some embodiments, the determination step, detection step, or sequence analysis involves contacting nucleic acid molecules in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe is i) adenine or its complement at the position corresponding to position 5,557 as described in SEQ ID NO: 3, ii) adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 22, or at position 617 as described in SEQ ID NO: 27 The method includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing adenine or its complement at a corresponding position, and / or iii) adenine or its complement at a position corresponding to position 556 as described in SEQ ID NO: 36, adenine or its complement at a position corresponding to position 457 as described in SEQ ID NO: 41, adenine or its complement at a position corresponding to position 378 as described in SEQ ID NO: 46, or adenine or its complement at a position corresponding to position 617 as described in SEQ ID NO: 51, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0143] In some embodiments, the determination step, detection step, or sequence analysis involves contacting a nucleic acid molecule in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe is i) thymine or its complement at the position corresponding to position 5,911 as described in SEQ ID NO: 4, ii) uracil or its complement at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, or at position 971 as described in SEQ ID NO: 28 The method includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing uracil or its complement at the corresponding position, and / or iii) thymine or its complement at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0144] In some embodiments, the determination step, detection step, or sequence analysis involves contacting nucleic acid molecules in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe is: i) lacking AAAG tetranucleotide or its complement at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1; ii) lacking AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 7; lacking AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 8; lacking AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 9; or position corresponding to positions 980-983 as described in SEQ ID NO: 10 The method includes contacting a nucleotide sequence of an amplified nucleic acid molecule that lacks an AAAG tetranucleotide or its complement, and / or iii) lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or lacks an AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0145] In some embodiments, the determination step, detection step, or sequence analysis involves contacting a nucleic acid molecule in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe i) inserts thymine or its complement at the position corresponding to position 6,411 as described in SEQ ID NO: 6, ii) inserts uracil or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 15, inserts uracil or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 20, inserts uracil or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 25, or at position 1,471 as described in SEQ ID NO: 30 The method includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing an insertion of uracil or its complement at a corresponding position, and / or iii) an insertion of thymine or its complement at the position corresponding to position 1,410 as described in SEQ ID NO: 39, an insertion of thymine or its complement at the position corresponding to position 1,311 as described in SEQ ID NO: 44, an insertion of thymine or its complement at the position corresponding to position 1,232 as described in SEQ ID NO: 49, or an insertion of thymine or its complement at the position corresponding to position 1,471 as described in SEQ ID NO: 54, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0146] In some embodiments, the determination step, detection step, or sequence analysis involves contacting a nucleic acid molecule in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe is i) guanine or its complement at the position corresponding to position 5,831 as described in SEQ ID NO: 99, ii) guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 101, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 102, or at position 891 as described in SEQ ID NO: 103 The method includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing guanine or its complement at a corresponding position, and / or iii) guanine or its complement at the position corresponding to position 830 as described in SEQ ID NO: 104, guanine or its complement at the position corresponding to position 731 as described in SEQ ID NO: 105, guanine or its complement at the position corresponding to position 652 as described in SEQ ID NO: 106, or guanine or its complement at the position corresponding to position 891 as described in SEQ ID NO: 107, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0147] In some embodiments, the determination step, detection step, or sequence analysis includes contacting a nucleic acid molecule in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe includes a nucleotide sequence that hybridizes under stringent conditions with the nucleotide sequence of an amplified nucleic acid molecule containing a nucleotide sequence that lacks a CCAGTAG heptanucleotide or its complement at positions 5,540-5,546 as described in SEQ ID NO: 2, contains adenine or its complement at position 5,557 as described in SEQ ID NO: 3, contains thymine or its complement at position 5,911 as described in SEQ ID NO: 4, lacks an AAAG tetranucleotide or its complement at positions 5,920-5,923 as described in SEQ ID NO: 5, contains an insertion of thymine or its complement at position 6,411 as described in SEQ ID NO: 6, or contains guanine or its complement at position 5,831 as described in SEQ ID NO: 99, and detecting the detectable label.

[0148] In some embodiments, the determination step, detection step, or sequence analysis involves contacting nucleic acid molecules in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe lacks CCAGUAG heptanucleotide or its complement at positions 539-545 as described in SEQ ID NO: 7, lacks CCAGUAG heptanucleotide or its complement at positions 440-446 as described in SEQ ID NO: 8, or lacks CCAGUAG heptanucleotide or its complement at positions 361-367 as described in SEQ ID NO: 9. The CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, which lacks its complement, or the adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, which lacks its complement, or the adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, which contains the adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, which contains the adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, which contains the adenine at the position corresponding to position 910 as described in SEQ ID NO: 13 Contains uracil or its complement, contains uracil or its complement at the position corresponding to position 811 as described in SEQ ID NO: 18, contains uracil or its complement at the position corresponding to position 732 as described in SEQ ID NO: 23, contains uracil or its complement at the position corresponding to position 971 as described in SEQ ID NO: 28, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 7, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 8, SEQ ID NO: 9 Lacking AAAG tetranucleotide or its complement at positions corresponding to 741-744 as described in SEQ ID NO: 10, Lacking AAAG tetranucleotide or its complement at positions corresponding to 980-983 as described in SEQ ID NO: 10, Including an insertion of uracil or its complement at position corresponding to 1,410 as described in SEQ ID NO: 15, Including an insertion of uracil or its complement at position corresponding to 1,311 as described in SEQ ID NO: 20, Including an insertion of uracil or its complement at position corresponding to 1,232 as described in SEQ ID NO: 25, 1,The process includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing an insertion of uracil or its complement at position 471, guanine or its complement at position 830 as described in SEQ ID NO: 100, guanine or its complement at position 731 as described in SEQ ID NO: 101, guanine or its complement at position 652 as described in SEQ ID NO: 102, or guanine or its complement at position 891 as described in SEQ ID NO: 103, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0149] In some embodiments, the determination step, detection step, or sequence analysis involves contacting nucleic acid molecules in a biological sample with a change-specific probe containing a detectable label, wherein the change-specific probe lacks a CCAGTAG heptanucleotide or its complement at positions 539-545 as described in SEQ ID NO: 31, a CCAGTAG heptanucleotide or its complement at positions 440-446 as described in SEQ ID NO: 32, or a CCAGTAG heptanucleotide at positions 361-367 as described in SEQ ID NO: 33. Lack of adenine or its complement, lacking a CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, lacking adenine or its complement, containing adenine or its complement at the position corresponding to position 556 as described in SEQ ID NO: 36, containing adenine or its complement at the position corresponding to position 457 as described in SEQ ID NO: 41, containing adenine or its complement at the position corresponding to position 378 as described in SEQ ID NO: 46, containing adenine or its complement at the position corresponding to position 617 as described in SEQ ID NO: 51, corresponding to position 910 as described in SEQ ID NO: 37 Contains thymine or its complement at the position, contains thymine or its complement at the position corresponding to position 811 as described in SEQ ID NO: 42, contains thymine or its complement at the position corresponding to position 732 as described in SEQ ID NO: 47, contains thymine or its complement at the position corresponding to position 971 as described in SEQ ID NO: 52, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in SEQ ID NO: 31, lacks AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in SEQ ID NO: 32, sequence number Lacking AAAG tetranucleotide or its complement at positions 741-744 as described in Sequence ID No. 33, Lacking AAAG tetranucleotide or its complement at positions 980-983 as described in Sequence ID No. 34, Includes insertion of thymine or its complement at position 1,410 as described in Sequence ID No. 39, Includes insertion of thymine or its complement at position 1,311 as described in Sequence ID No. 44, Includes insertion of thymine or its complement at position 1,232 as described in Sequence ID No. 49, 1,The process includes contacting a nucleotide sequence of an amplified nucleic acid molecule containing an insertion of thymine or its complement at position 471, guanine or its complement at position 830 as described in SEQ ID NO: 104, guanine or its complement at position 731 as described in SEQ ID NO: 105, guanine or its complement at position 652 as described in SEQ ID NO: 106, or guanine or its complement at position 891 as described in SEQ ID NO: 107, with a nucleotide sequence that hybridizes under stringent conditions, and detecting a detectable label.

[0150] Mutation-specific polymerase chain reaction (SMP) technology can be used to detect mutations such as SNPs in nucleic acid sequences. Since DNA polymerase will not extend if a mismatch with the template is present, mutation-specific primers can be used.

[0151] In some embodiments, the nucleic acid molecule in the sample is mRNA, and the mRNA is reverse transcribed to cDNA before the amplification step. In some embodiments, the nucleic acid molecule is present in cells obtained from the subject.

[0152] In some embodiments, the assay includes contacting a biological sample with a primer or probe, such as a variant-specific primer or probe, that specifically hybridizes with a GPR75 variant genome sequence, variant mRNA sequence, or variant cDNA sequence under stringent conditions but does not hybridize with the corresponding GPR75 reference sequence, and determining whether hybridization has occurred.

[0153] In some embodiments, the assay includes RNA sequencing (RNA-Seq). In some embodiments, the assay also includes reverse transcription of mRNA into cDNA, for example, by reverse transcription polymerase chain reaction (RT-PCR).

[0154] In some embodiments, the method utilizes probes and primers of sufficient nucleotide length to bind to a target nucleotide sequence and specifically detect and / or identify polynucleotides, including GPR75 variant genomic nucleic acid molecules, variant mRNA molecules, or variant cDNA molecules. Hybridization or reaction conditions can be determined by the operator to achieve this result. This nucleotide length may be any length sufficient for use in selected detection methods, including any assay described or exemplified herein. Such probes and primers can specifically hybridize to the target nucleotide sequence under high stringency hybridization conditions. While probes and primers may have complete nucleotide sequence identity of consecutive nucleotides within the target nucleotide sequence, probes can be conventionally designed to be distinct from the target nucleotide sequence while retaining the ability to specifically detect and / or identify the target nucleotide sequence. The probes and primers may have approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity with respect to the nucleotide sequence of the target nucleic acid molecule.

[0155] In some embodiments, the GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in the biological sample is a nucleotide sequence (genomic nucleic acid molecule) lacking the CCAGUAG heptanucleotide at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, a deletion of the CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, or a deletion of the C at the position corresponding to positions 361-367 as described in SEQ ID NO: 9. Deletion of CAGUAG heptanucleotide, or deletion of CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10 (in the case of mRNA molecules); deletion of CCAGTAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, deletion of CCAGTAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, deletion of CCAGTAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or CCAG at the position corresponding to positions 600-606 as described in SEQ ID NO: 34 To determine whether or not a TAG heptanucleotide deletion (in the case of a cDNA molecule) is present, the following deletions are identified: the CCAGUAG heptanucleotide deletion at the position corresponding to positions 5,540-5,546 as described in SEQ ID NO: 1, the CCAGUAG heptanucleotide deletion at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, the CCAGUAG heptanucleotide deletion at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, the CCAGUAG heptanucleotide deletion at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, and the position corresponding to positions 600-606 as described in SEQ ID NO: 10. A first primer derived from the 5' adjacent sequence adjacent to the CCAGUAG heptanucleotide deletion, the CCAGTAG heptanucleotide deletion at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, the CCAGTAG heptanucleotide deletion at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, the CCAGTAG heptanucleotide deletion at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or the CCAGTAG heptanucleotide deletion at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, and the 5,540-5,Deletion of the CCAGUAG heptanucleotide at position 546, deletion of the CCAGUAG heptanucleotide at positions 539-545 as described in SEQ ID NO: 7, deletion of the CCAGUAG heptanucleotide at positions 440-446 as described in SEQ ID NO: 8, deletion of the CCAGUAG heptanucleotide at positions 361-367 as described in SEQ ID NO: 9, deletion of the CCAGUAG heptanucleotide at positions 600-606 as described in SEQ ID NO: 10, and deletion of the CCAGUAG heptanucleotide at positions 539-545 as described in SEQ ID NO: 31 A biological sample is subjected to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to a deletion of a CCAGTAG heptanucleotide at the specified position, a deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, a deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or a deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, and the 5,540-5,It is possible to generate amplicons that show the presence of a deletion of CCAGUAG heptanucleotide at the position corresponding to position 546, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 7, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 8, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 9, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 10, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 539-545 as described in SEQ ID NO: 31, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or a deletion of CCAGUAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34. In some embodiments, the amplicon length may range from the combined length of one nucleotide base pair of the primer pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the amplification limit, or approximately 20,000 nucleotide base pairs. Optionally, the primer pair may be 5,540~5, as described in Sequence ID No. 1.The position where the CCAGUAG heptanucleotide is missing at position 546, the deletion of the CCAGUAG heptanucleotide at positions 539-545 as described in SEQ ID NO: 7, the deletion of the CCAGUAG heptanucleotide at positions 440-446 as described in SEQ ID NO: 8, the deletion of the CCAGUAG heptanucleotide at positions 361-367 as described in SEQ ID NO: 9, the deletion of the CCAGUAG heptanucleotide at positions 600-606 as described in SEQ ID NO: 10, sequence number Deletion of CCAGTAG heptanucleotide at positions 539-545 as described in SEQ ID NO. 31, deletion of CCAGTAG heptanucleotide at positions 440-446 as described in SEQ ID NO. 32, deletion of CCAGTAG heptanucleotide at positions 361-367 as described in SEQ ID NO. 33, or deletion of CCAGTAG heptanucleotide at positions 600-606 as described in SEQ ID NO. 34, and CCAGTAG at positions 5,540-5,546 as described in SEQ ID NO. 1 The following are examples of missing heptanucleotides: deletion of CCAGUAG heptanucleotides at positions 539-545 as described in SEQ ID NO: 7, deletion of CCAGUAG heptanucleotides at positions 440-446 as described in SEQ ID NO: 8, deletion of CCAGUAG heptanucleotides at positions 361-367 as described in SEQ ID NO: 9, deletion of CCAGUAG heptanucleotides at positions 600-606 as described in SEQ ID NO: 10, and deletion of CCAGUAG heptanucleotides at positions 539-545 as described in SEQ ID NO: 31. The region is adjacent to at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more nucleotides on each side of the deletion of a CCAGTAG heptanucleotide at the specified position, the deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 440-446 as described in SEQ ID NO: 32, the deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 361-367 as described in SEQ ID NO: 33, or the deletion of a CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in SEQ ID NO: 34, and the region containing these nucleotides.

[0156] In some embodiments, the GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in the biological sample is adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3 (genomic nucleic acid molecule), adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 27 (in the case of an mRNA molecule); at position 556 as described in SEQ ID NO: 36 To determine whether a nucleotide sequence contains an adenine at the corresponding position, an adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, an adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or an adenine at the position corresponding to position 617 as described in SEQ ID NO: 51 (in the case of a cDNA molecule), the adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3, an adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, an adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, an adenine at the position corresponding to position 378 as described in SEQ ID NO: 22 A first primer derived from a 5' adjacent sequence adjacent to adenine, the adenine at position 617 as described in SEQ ID NO: 27, the adenine at position 556 as described in SEQ ID NO: 36, the adenine at position 457 as described in SEQ ID NO: 41, the adenine at position 378 as described in SEQ ID NO: 46, or the adenine at position 617 as described in SEQ ID NO: 51, and the adenine at position 5,557 as described in SEQ ID NO: 3, the adenine at position 556 as described in SEQ ID NO: 12, and the adenine at position 457 as described in SEQ ID NO: 17 A biological sample is subjected to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to the adenine at the position described in SEQ ID NO: 3, the adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, the adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, the adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, the adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, the adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or the adenine at the position corresponding to position 617 as described in SEQ ID NO: 51, and the 5, described in SEQ ID NO: 3.It is possible to generate amplicons that indicate the presence of an SNP at the position encoding the adenine at position 557, the adenine at position 556 as described in SEQ ID NO: 12, the adenine at position 457 as described in SEQ ID NO: 17, the adenine at position 378 as described in SEQ ID NO: 22, the adenine at position 617 as described in SEQ ID NO: 27, the adenine at position 556 as described in SEQ ID NO: 36, the adenine at position 457 as described in SEQ ID NO: 41, the adenine at position 378 as described in SEQ ID NO: 46, or the adenine at position 617 as described in SEQ ID NO: 51. In some embodiments, the length of the amplicon may range from the length of one nucleotide base pair combined with the primer pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the limit of the amplification reaction, or to about 20,000 nucleotide base pairs. Optionally, the primer pair may include a position containing adenine at the position corresponding to position 5,557 as described in SEQ ID NO: 3, adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or adenine at the position corresponding to position 617 as described in SEQ ID NO: 51, and a position corresponding to position 5,557 as described in SEQ ID NO: 3 The region is adjacent to a region containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the position containing the adenine at the position, the adenine at the position corresponding to position 556 as described in SEQ ID NO: 12, the adenine at the position corresponding to position 457 as described in SEQ ID NO: 17, the adenine at the position corresponding to position 378 as described in SEQ ID NO: 22, the adenine at the position corresponding to position 617 as described in SEQ ID NO: 27, the adenine at the position corresponding to position 556 as described in SEQ ID NO: 36, the adenine at the position corresponding to position 457 as described in SEQ ID NO: 41, the adenine at the position corresponding to position 378 as described in SEQ ID NO: 46, or the adenine at the position corresponding to position 617 as described in SEQ ID NO: 51.

[0157] In some embodiments, to determine whether a GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in a biological sample contains a nucleotide sequence including thymine at the position corresponding to position 5,911 as described in SEQ ID NO: 4 (genomic nucleic acid molecule), uracil at the position corresponding to position 910 as described in SEQ ID NO: 13, uracil at the position corresponding to position 811 as described in SEQ ID NO: 18, uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, or uracil at the position corresponding to position 971 as described in SEQ ID NO: 28 (in the case of an mRNA molecule); thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or thymine at the position corresponding to position 971 as described in SEQ ID NO: 52 (in the case of a cDNA molecule), the following are used: A first primer derived from a 5' adjacent sequence adjacent to uracil, uracil at position 971 as described in SEQ ID NO: 28, thymine at position 910 as described in SEQ ID NO: 37, thymine at position 811 as described in SEQ ID NO: 42, thymine at position 732 as described in SEQ ID NO: 47, or thymine at position 971 as described in SEQ ID NO: 52, and thymine at position 5,911 as described in SEQ ID NO: 4, uracil at position 910 as described in SEQ ID NO: 13, and position 811 as described in SEQ ID NO: 18 A biological sample is subjected to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to the uracil at the corresponding position, the uracil at the position corresponding to position 732 as described in SEQ ID NO: 23, the uracil at the position corresponding to position 971 as described in SEQ ID NO: 28, the thymine at the position corresponding to position 910 as described in SEQ ID NO: 37, the thymine at the position corresponding to position 811 as described in SEQ ID NO: 42, the thymine at the position corresponding to position 732 as described in SEQ ID NO: 47, or the thymine at the position corresponding to position 971 as described in SEQ ID NO: 52, as described in SEQ ID NO: 4.An amplicon can be generated that indicates the presence of an SNP at the position encoding thymine at position 911, uracil at position 910 as described in SEQ ID NO: 13, uracil at position 811 as described in SEQ ID NO: 18, uracil at position 732 as described in SEQ ID NO: 23, uracil at position 971 as described in SEQ ID NO: 28, thymine at position 910 as described in SEQ ID NO: 37, thymine at position 811 as described in SEQ ID NO: 42, thymine at position 732 as described in SEQ ID NO: 47, or thymine at position 971 as described in SEQ ID NO: 52. In some embodiments, the length of the amplicon may range from the length of one nucleotide base pair combined with the primer pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the limit of the amplification reaction, or about 20,000 nucleotide base pairs. Optionally, the primer pair includes a position containing thymine at position 5,911 as described in SEQ ID NO: 4, uracil at position 910 as described in SEQ ID NO: 13, uracil at position 811 as described in SEQ ID NO: 18, uracil at position 732 as described in SEQ ID NO: 23, uracil at position 971 as described in SEQ ID NO: 28, thymine at position 910 as described in SEQ ID NO: 37, thymine at position 811 as described in SEQ ID NO: 42, thymine at position 732 as described in SEQ ID NO: 47, or thymine at position 971 as described in SEQ ID NO: 52, and the position corresponding to position 5,911 as described in SEQ ID NO: 4 The region is adjacent to a region containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the position containing thymine at position 910, uracil at position 910 as described in SEQ ID NO: 13, uracil at position 811 as described in SEQ ID NO: 18, uracil at position 732 as described in SEQ ID NO: 23, uracil at position 971 as described in SEQ ID NO: 28, thymine at position 910 as described in SEQ ID NO: 37, thymine at position 811 as described in SEQ ID NO: 42, thymine at position 732 as described in SEQ ID NO: 47, or thymine at position 971 as described in SEQ ID NO: 52.

[0158] In some embodiments, the GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in the biological sample lacks an AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 as described in SEQ ID NO: 1, a deletion of an AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, a deletion of an AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, a deletion of an AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, or a deletion of an AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10 (in the case of mRNA molecules); a deletion of an AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, a deletion of an AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, a deletion of an AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or an A at the position corresponding to positions 980-983 as described in SEQ ID NO: 34 To determine whether or not a cDNA molecule contains an AAG tetranucleotide deletion, we examine the following: the deletion of AAAG tetranucleotide at positions 5,920-5,923 as described in SEQ ID NO: 1, the deletion of AAAG tetranucleotide at positions 919-922 as described in SEQ ID NO: 7, the deletion of AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 8, the deletion of AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 9, and the deletion at positions 980-983 as described in SEQ ID NO: 10. A first primer derived from the 5' adjacent sequence adjacent to the deletion of AAAG tetranucleotide at the position, the deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, the deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, the deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or the deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, and the 5,920-5,A biological sample is subjected to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to the deletion of AAAG tetranucleotide at the position corresponding to position 923, the deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, the deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, the deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, the deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 10, the deletion of AAAG tetranucleotide at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, the deletion of AAAG tetranucleotide at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, the deletion of AAAG tetranucleotide at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or the deletion of AAAG tetranucleotide at the position corresponding to positions 980-983 as described in SEQ ID NO: 34, and the 5,920-5,It is possible to generate amplicons showing the presence of an AAAG tetranucleotide deletion at the position corresponding to position 923, an AAAG tetranucleotide deletion at the position corresponding to positions 919-922 as described in SEQ ID NO: 7, an AAAG tetranucleotide deletion at the position corresponding to positions 820-823 as described in SEQ ID NO: 8, an AAAG tetranucleotide deletion at the position corresponding to positions 741-744 as described in SEQ ID NO: 9, an AAAG tetranucleotide deletion at the position corresponding to positions 980-983 as described in SEQ ID NO: 10, an AAAG tetranucleotide deletion at the position corresponding to positions 919-922 as described in SEQ ID NO: 31, an AAAG tetranucleotide deletion at the position corresponding to positions 820-823 as described in SEQ ID NO: 32, an AAAG tetranucleotide deletion at the position corresponding to positions 741-744 as described in SEQ ID NO: 33, or an AAAG tetranucleotide deletion at the position corresponding to positions 980-983 as described in SEQ ID NO: 34. In some embodiments, the length of the amplicon may range from the length of a primer pair combined with one nucleotide base pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the amplification limit, or approximately 20,000 nucleotide base pairs. Optionally, the primer pair may be a position lacking AAAG tetranucleotide at positions 5,920-5,923 as described in SEQ ID NO: 1, a deletion of AAAG tetranucleotide at positions 919-922 as described in SEQ ID NO: 7, a deletion of AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 8, a deletion of AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 9, a deletion of AAAG tetranucleotide at positions 980-983 as described in SEQ ID NO: 10, a deletion of AAAG tetranucleotide at positions 919-922 as described in SEQ ID NO: 31, a deletion of AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 32, a deletion of AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 33, or a deletion of AAAG tetranucleotide at positions 980-983 as described in SEQ ID NO: 34, and a position lacking AAAG tetranucleotide at positions 5,920-5,The region is adjacent to the region containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the following locations: the position lacking the AAAG tetranucleotide at position 923, the deletion of the AAAG tetranucleotide at positions 919-922 as described in SEQ ID NO: 7, the deletion of the AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 8, the deletion of the AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 9, the deletion of the AAAG tetranucleotide at positions 980-983 as described in SEQ ID NO: 10, the deletion of the AAAG tetranucleotide at positions 919-922 as described in SEQ ID NO: 31, the deletion of the AAAG tetranucleotide at positions 820-823 as described in SEQ ID NO: 32, the deletion of the AAAG tetranucleotide at positions 741-744 as described in SEQ ID NO: 33, or the deletion of the AAAG tetranucleotide at positions 980-983 as described in SEQ ID NO: 34.

[0159] In some embodiments, the GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in the biological sample is an insertion of thymine at the position corresponding to position 6,411 as described in SEQ ID NO: 6 (genomic nucleic acid molecule), an insertion of uracil at the position corresponding to position 1,410 as described in SEQ ID NO: 15, an insertion of uracil at the position corresponding to position 1,311 as described in SEQ ID NO: 20, an insertion of uracil at the position corresponding to position 1,232 as described in SEQ ID NO: 25, or an insertion of uracil at the position corresponding to position 1,471 as described in SEQ ID NO: 30 (in the case of mRNA molecule); To determine whether a nucleotide sequence contains a thymine insertion at the position corresponding to position 1,410 as described in sequence number 39, a thymine insertion at the position corresponding to position 1,311 as described in sequence number 44, a thymine insertion at the position corresponding to position 1,232 as described in sequence number 49, or a thymine insertion at the position corresponding to position 1,471 as described in sequence number 54 (in the case of a cDNA molecule), the thymine insertion at the position corresponding to position 6,411 as described in sequence number 6, a uracil insertion at the position corresponding to position 1,410 as described in sequence number 15, and the position corresponding to position 1,311 as described in sequence number 20 A first primer derived from a 5' adjacency sequence adjacent to the uracil insertion described in SEQ ID NO: 6, the uracil insertion at position 1,232 as described in SEQ ID NO: 25, the uracil insertion at position 1,471 as described in SEQ ID NO: 30, the thymine insertion at position 1,410 as described in SEQ ID NO: 39, the thymine insertion at position 1,311 as described in SEQ ID NO: 44, the thymine insertion at position 1,232 as described in SEQ ID NO: 49, or the thymine insertion at position 1,471 as described in SEQ ID NO: 54, and the uracil insertion at position 6,411 as described in SEQ ID NO: 6 Insertion of thymine at position, insertion of uracil at position 1,410 as described in SEQ ID NO: 15, insertion of uracil at position 1,311 as described in SEQ ID NO: 20, insertion of uracil at position 1,232 as described in SEQ ID NO: 25, insertion of uracil at position 1,471 as described in SEQ ID NO: 30, insertion of thymine at position 1,410 as described in SEQ ID NO: 39, insertion of thymine at position 1,311 as described in SEQ ID NO: 44, insertion of thymine at position 1,232 as described in SEQ ID NO: 49, or 1,By subjecting a biological sample to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to the thymine insertion at position 471, it is possible to generate an amplicon showing the presence of the thymine insertion at position 6,411 as described in SEQ ID NO: 6, the uracil insertion at position 1,410 as described in SEQ ID NO: 15, the uracil insertion at position 1,311 as described in SEQ ID NO: 20, the uracil insertion at position 1,232 as described in SEQ ID NO: 25, the uracil insertion at position 1,471 as described in SEQ ID NO: 30, the thymine insertion at position 1,410 as described in SEQ ID NO: 39, the thymine insertion at position 1,311 as described in SEQ ID NO: 44, the thymine insertion at position 1,232 as described in SEQ ID NO: 49, or the thymine insertion at position 1,471 as described in SEQ ID NO: 54. In some embodiments, the amplicon length may range from the combined length of one nucleotide base pair of the primer pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the amplification limit, or approximately 20,000 nucleotide base pairs. Optionally, the primer pair may be: a thymine insertion at position 6,411 as described in SEQ ID NO: 6, a uracil insertion at position 1,410 as described in SEQ ID NO: 15, a uracil insertion at position 1,311 as described in SEQ ID NO: 20, a uracil insertion at position 1,232 as described in SEQ ID NO: 25, a uracil insertion at position 1,471 as described in SEQ ID NO: 30, a thymine insertion at position 1,410 as described in SEQ ID NO: 39, a thymine insertion at position 1,311 as described in SEQ ID NO: 44, a sequence number The insertion of thymine at position 1,232 as described in Sequence No. 49, or the insertion of thymine at position 1,471 as described in Sequence No. 54, and the insertion of thymine at position 6,411 as described in Sequence No. 6, the insertion of uracil at position 1,410 as described in Sequence No. 15, the insertion of uracil at position 1,311 as described in Sequence No. 20, the insertion of uracil at position 1,232 as described in Sequence No. 25, the insertion of uracil at position 1,471 as described in Sequence No. 30, 1,The region is adjacent to a region containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the position including the thymine insertion at position 410, the thymine insertion at position 1,311 as described in SEQ ID NO: 44, the thymine insertion at position 1,232 as described in SEQ ID NO: 49, or the thymine insertion at position 1,471 as described in SEQ ID NO: 54.

[0160] In some embodiments, the GPR75 nucleic acid molecule (genomic nucleic acid molecule, mRNA molecule, or cDNA molecule) or its complement in the biological sample is guanine at the position corresponding to position 5,831 as described in SEQ ID NO: 99 (genomic nucleic acid molecule), guanine at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine at the position corresponding to position 731 as described in SEQ ID NO: 101, guanine at the position corresponding to position 652 as described in SEQ ID NO: 102, or guanine at the position corresponding to position 891 as described in SEQ ID NO: 103 (in the case of an mRNA molecule); corresponding to position 830 as described in SEQ ID NO: 104 To determine whether a nucleotide sequence contains guanine at the corresponding position, guanine at the position corresponding to position 731 as described in SEQ ID NO: 105, guanine at the position corresponding to position 652 as described in SEQ ID NO: 106, or guanine at the position corresponding to position 891 as described in SEQ ID NO: 107 (in the case of a cDNA molecule), the nucleotide sequence contains guanine at the position corresponding to position 5,831 as described in SEQ ID NO: 99, guanine at the position corresponding to position 830 as described in SEQ ID NO: 100, guanine at the position corresponding to position 731 as described in SEQ ID NO: 101, and guanine at the position corresponding to position 652 as described in SEQ ID NO: 102. A first primer derived from the 5' adjacent sequence adjacent to the guanine at position 891 described in SEQ ID NO: 103, the guanine at position 830 described in SEQ ID NO: 104, the guanine at position 731 described in SEQ ID NO: 105, the guanine at position 652 described in SEQ ID NO: 106, or the guanine at position 891 described in SEQ ID NO: 107, and the guanine at position 5,831 described in SEQ ID NO: 99, the guanine at position 830 described in SEQ ID NO: 100, and the guanine at position 731 described in SEQ ID NO: 101 A biological sample is subjected to an amplification method using a primer pair comprising a second primer derived from the 3' adjacent sequence adjacent to the guanine at the position described in SEQ ID NO: 102, the guanine at the position corresponding to position 652 as described in SEQ ID NO: 103, the guanine at the position corresponding to position 830 as described in SEQ ID NO: 104, the guanine at the position corresponding to position 731 as described in SEQ ID NO: 105, the guanine at the position corresponding to position 652 as described in SEQ ID NO: 106, or the guanine at the position corresponding to position 891 as described in SEQ ID NO: 107, and the 5, described in SEQ ID NO: 99.An amplicon can be generated that shows the presence of guanine insertion at position 831, guanine at position 830 as described in SEQ ID NO: 100, guanine at position 731 as described in SEQ ID NO: 101, guanine at position 652 as described in SEQ ID NO: 102, guanine at position 891 as described in SEQ ID NO: 103, guanine at position 830 as described in SEQ ID NO: 104, guanine at position 731 as described in SEQ ID NO: 105, guanine at position 652 as described in SEQ ID NO: 106, or guanine at position 891 as described in SEQ ID NO: 107. In some embodiments, the length of the amplicon may range from the length of one nucleotide base pair combined with the primer pair to any length of amplicon that can be generated by the DNA amplification protocol. This distance can range from one nucleotide base pair to the limit of the amplification reaction, or to about 20,000 nucleotide base pairs. Optionally, the primer pair may include a guanine insertion at the position corresponding to position 5,831 as described in SEQ ID NO: 99, a guanine at the position corresponding to position 830 as described in SEQ ID NO: 100, a guanine at the position corresponding to position 731 as described in SEQ ID NO: 101, a guanine at the position corresponding to position 652 as described in SEQ ID NO: 102, a guanine at the position corresponding to position 891 as described in SEQ ID NO: 103, a guanine at the position corresponding to position 830 as described in SEQ ID NO: 104, a guanine at the position corresponding to position 731 as described in SEQ ID NO: 105, a guanine at the position corresponding to position 652 as described in SEQ ID NO: 106, or a guanine at the position corresponding to position 891 as described in SEQ ID NO: 107, and position 5,The region is adjacent to a region containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the position containing the guanine at position 831, the guanine at position 830 as described in SEQ ID NO: 100, the guanine at position 731 as described in SEQ ID NO: 101, the guanine at position 652 as described in SEQ ID NO: 102, the guanine at position 891 as described in SEQ ID NO: 103, the guanine at position 830 as described in SEQ ID NO: 104, the guanine at position 731 as described in SEQ ID NO: 105, the guanine at position 652 as described in SEQ ID NO: 106, or the guanine at position 891 as described in SEQ ID NO: 107.

[0161] Similar amplicons can be constructed from mRNA and / or cDNA sequences. PCR primer pairs can be derived from known sequences using computer programs intended for this purpose, such as the PCR primer analysis tools in Vector NTI version 10 (Informax Inc., Bethesda Md.), PrimerSelect (DNASTAR Inc., Madison, Wis.), and Primer3 (version 0.4.0.COPYRGT., 1991, Whitehead Institute for Biomedical Research, Cambridge, Mass.). In addition, sequences can be visually inspected and primers manually identified using known guidelines.

[0162] Examples of nucleic acid sequencing techniques include, but are not limited to, chain arrest (Sanger) sequencing and diterminator sequencing. Other methods include non-sequencing nucleic acid hybridization methods, including the use of labeled primers or probes on purified DNA, amplified DNA, and fixed cell specimens (fluorescence in situ hybridization (FISH)). In some methods, target nucleic acid molecules can be amplified before detection or simultaneously with detection. Examples of nucleic acid amplification techniques include, but are not limited to, polymerase chain reaction (PCR), ligase chain reaction (LCR), strand displacement amplification (SDA), and nucleic acid sequence-based amplification (NASBA). Other methods include, but are not limited to, ligase chain reaction, strand displacement amplification, and high-temperature SDA (tSDA).

[0163] In hybridization techniques, stringent conditions may be used to ensure that the probe or primer specifically hybridizes with its target. In some embodiments, under stringent conditions, a polynucleotide primer or probe hybridizes with its target sequence to a degree that is more detectable than other non-target sequences, for example, at least twice, at least three times, at least four times, or more (including more than ten times the background). In some embodiments, a polynucleotide primer or probe hybridizes with its target nucleotide sequence to a degree that is at least twice as detectable as other nucleotide sequences. In some embodiments, a polynucleotide primer or probe hybridizes with its target nucleotide sequence to a degree that is at least three times as detectable as other nucleotide sequences. In some embodiments, a polynucleotide primer or probe hybridizes with its target nucleotide sequence to a degree that is at least four times as detectable as other nucleotide sequences. In some embodiments, under stringent conditions, a polynucleotide primer or probe hybridizes with its target nucleotide sequence to a degree that is more detectable than other nucleotide sequences (more than 10 times higher than background). Stringent conditions are sequence-dependent and vary depending on the context.

[0164] Appropriate stringing conditions to promote DNA hybridization, such as washing with 6× sodium chloride / sodium citrate (SSC) at approximately 45°C followed by 2× SSC at 50°C, are known or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, NY (1989), 6.3.1-6.3.6. Typically, stringing conditions for hybridization and detection involve a salt concentration of Na at pH 7.0-8.3. + Ions less than approximately 1.5M, typically Na+ The conditions are that the ion concentration (or other salt) is approximately 0.01–1.0 M, and the temperature is at least approximately 30°C for short probes (e.g., 10–50 nucleotides) and at least approximately 60°C for longer probes (e.g., over 50 nucleotides). Stringent conditions can also be achieved by adding a destabilizing agent such as formamide. Optionally, the wash buffer may contain approximately 0.1%–1% SDS. The duration of hybridization is generally less than approximately 24 hours, usually about 4–12 hours. The duration of washing should be at least long enough to reach equilibrium.

[0165] This disclosure also provides a method for detecting the presence of a predicted loss-of-function polypeptide of human GPR75, comprising performing an assay on a biological sample obtained from a subject to determine whether the GPR75 polypeptide of the subject contains one or more mutations that cause the polypeptide to have loss-of-function (partial or complete) or predicted loss-of-function (partial or complete). The predicted loss-of-function polypeptide of GPR75 may be any of the GPR75 variant polypeptides described herein. In some embodiments, the method detects the presence of GPR75 Ala110fs, Ala116Thr, Tyr207Cys, Gln234Stop, Arg236fs, or Cys400fs. In some embodiments, the method detects Lys404 * Detects the presence of Ser219fs or c.-110+1G>A.

[0166] In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample contains amino acids 110-130 as described in SEQ ID NO: 56. In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample contains threonine at the position corresponding to position 116 as described in SEQ ID NO: 57. In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample terminates at the position corresponding to position 233 as described in SEQ ID NO: 58. In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample contains amino acids 236-239 as described in SEQ ID NO: 59. In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample contains amino acids 400-425 as described in SEQ ID NO: 60. In some embodiments, the method includes performing an assay on a sample obtained from a subject to determine whether the GPR75 polypeptide in the sample contains cysteine ​​at the position corresponding to position 207 as described in SEQ ID NO: 108.

[0167] In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to one or more of the positions 110-130 described in SEQ ID NO: 56 or SEQ ID NO: 55. In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to the position 116 described in SEQ ID NO: 57 or SEQ ID NO: 55. In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to one or more of the positions 234-540 described in SEQ ID NO: 55. In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to one or more of the positions 236-239 described in SEQ ID NO: 59 or SEQ ID NO: 55. In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to one or more of the positions 400-425 described in SEQ ID NO: 60 or SEQ ID NO: 55. In some embodiments, the detection step includes sequencing at least a portion of the polypeptide containing a position corresponding to the position 207 described in SEQ ID NO: 108.

[0168] In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to one or more of the 110-130 positions described in SEQ ID NO: 56 or SEQ ID NO: 55. In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to the 116 position described in SEQ ID NO: 57 or SEQ ID NO: 55. In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to one or more of the 234-540 positions described in SEQ ID NO: 55. In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to one or more of the 236-239 positions described in SEQ ID NO: 59 or SEQ ID NO: 55. In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to one or more of the 400-425 positions described in SEQ ID NO: 60 or SEQ ID NO: 55. In some embodiments, the detection step includes an immunoassay for detecting the presence of a polypeptide containing a position corresponding to the 207 position described in SEQ ID NO: 108 or SEQ ID NO: 55.

[0169] In some embodiments, if a subject does not have the predicted loss-of-function polypeptide of GPR75, the subject is at high risk of developing obesity or overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake. In some embodiments, if a subject has the predicted loss-of-function polypeptide of GPR75, the subject is at low risk of developing obesity or overweight, elevated BMI, elevated body fat mass, body fat percentage, or body fat volume, and / or excessive food intake.

[0170] This disclosure also provides isolated nucleic acid molecules that hybridize with GPR75 variant genomic nucleic acid molecules, GPR75 variant mRNA molecules, and / or GPR75 variant cDNA molecules (e.g., any of the genomic variant nucleic acid molecules, mRNA variant molecules, and cDNA variant molecules disclosed herein). In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes positions corresponding to positions 5,540-5,546 as described in SEQ ID NO: 2, positions 539-545 as described in SEQ ID NO: 11, positions 440-446 as described in SEQ ID NO: 16, positions 361-367 as described in SEQ ID NO: 21, positions 600-606 as described in SEQ ID NO: 26, positions 539-545 as described in SEQ ID NO: 35, positions 440-446 as described in SEQ ID NO: 40, positions 361-367 as described in SEQ ID NO: 45, or positions 600-606 as described in SEQ ID NO: 50.

[0171] In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes a position corresponding to position 5,557 as described in SEQ ID NO: 3, position 556 as described in SEQ ID NO: 12, position 457 as described in SEQ ID NO: 17, position 378 as described in SEQ ID NO: 22, position 617 as described in SEQ ID NO: 27, position 556 as described in SEQ ID NO: 36, position 457 as described in SEQ ID NO: 41, position 378 as described in SEQ ID NO: 46, or position 617 as described in SEQ ID NO: 51.

[0172] In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes a position corresponding to position 5,911 as described in SEQ ID NO: 4, position 910 as described in SEQ ID NO: 13, position 811 as described in SEQ ID NO: 18, position 732 as described in SEQ ID NO: 23, position 971 as described in SEQ ID NO: 28, position 910 as described in SEQ ID NO: 37, position 811 as described in SEQ ID NO: 42, position 732 as described in SEQ ID NO: 47, or position 971 as described in SEQ ID NO: 52.

[0173] In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes positions corresponding to positions 5,920-5,923 described in SEQ ID NO: 5, 919-922 described in SEQ ID NO: 14, 820-823 described in SEQ ID NO: 19, 741-744 described in SEQ ID NO: 24, 980-983 described in SEQ ID NO: 29, 919-922 described in SEQ ID NO: 38, 820-823 described in SEQ ID NO: 43, 741-744 described in SEQ ID NO: 48, or 980-983 described in SEQ ID NO: 53.

[0174] In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes a position corresponding to position 6,411 as described in SEQ ID NO: 6, position 1,410 as described in SEQ ID NO: 15, position 1,311 as described in SEQ ID NO: 20, position 1,232 as described in SEQ ID NO: 25, position 1,471 as described in SEQ ID NO: 30, position 1,410 as described in SEQ ID NO: 39, position 1,311 as described in SEQ ID NO: 44, position 1,232 as described in SEQ ID NO: 49, or position 1,471 as described in SEQ ID NO: 54.

[0175] In some embodiments, the isolated nucleic acid molecule hybridizes with a portion of the GPR75 nucleic acid molecule that includes a position corresponding to position 5,831 as described in SEQ ID NO: 99, position 830 as described in SEQ ID NO: 100, position 731 as described in SEQ ID NO: 101, position 652 as described in SEQ ID NO: 102, position 891 as described in SEQ ID NO: 103, position 830 as described in SEQ ID NO: 104, position 731 as described in SEQ ID NO: 105, position 652 as described in SEQ ID NO: 106, or position 891 as described in SEQ ID NO: 107.

[0176] For several purposes, such isolated nucleic acid molecules are at least about 5, at least about 8, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 5 5, comprising or consisting of at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, at least about 3000, at least about 4000, or at least about 5000 nucleotides. In some embodiments, such isolated nucleic acid molecules comprising or consisting of at least about 5, at least about 8, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, or at least about 25 nucleotides. In some embodiments, the isolated nucleic acid molecule contains or comprises at least about 18 nucleotides. In some embodiments, the isolated nucleic acid molecule contains or comprises at least about 15 nucleotides. In some embodiments, the isolated nucleic acid molecule contains or comprises about 10 to about 35, about 10 to about 30, about 10 to about 25, about 12 to about 30, about 12 to about 28, about 12 to about 24, about 15 to about 30, about 15 to about 25, about 18 to about 30, about 18 to about 25, about 18 to about 24, or about 18 to about 22 nucleotides. In some embodiments, the isolated nucleic acid molecule contains or comprises about 18 to about 30 nucleotides.In some embodiments, the isolated nucleic acid molecule contains or consists of at least about 15 nucleotides to at least about 35 nucleotides.

[0177] In some embodiments, such isolated nucleic acid molecules hybridize with GPR75 variant nucleic acid molecules (e.g., genomic nucleic acid molecules, mRNA molecules, and / or cDNA molecules) under stringent conditions. Such nucleic acid molecules can be used, for example, as probes, primers, mutation-specific probes, or mutation-specific primers as described or illustrated herein, including, but not limited to, primers, probes, antisense RNA, shRNA, and siRNA, each of which is detailed elsewhere herein, and which can be used in any of the methods described herein.

[0178] In some embodiments, the isolated nucleic acid molecule hybridizes with at least about 15 consecutive nucleotides of a nucleic acid molecule that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to the GPR75 variant genomic nucleic acid molecule, the GPR75 variant mRNA molecule, and / or the GPR75 variant cDNA molecule. In some embodiments, the isolated nucleic acid molecule consists of or contains about 15 to about 100 nucleotides, or about 15 to about 35 nucleotides. In some embodiments, the isolated nucleic acid molecule consists of or contains about 15 to about 100 nucleotides. In some embodiments, the isolated nucleic acid molecule consists of or contains about 15 to about 35 nucleotides.

[0179] In some embodiments, the isolated change-specific probe or change-specific primer comprises at least about 15 nucleotides, the change-specific probe or change-specific primer comprising a nucleotide sequence complementary to a portion of the nucleotide sequence encoding the human GPR75 polypeptide, the portion comprising the position corresponding to or its complement at positions 5,540-5,546 as described in SEQ ID NO: 2, the position corresponding to or its complement at positions 539-545 as described in SEQ ID NO: 11, the position corresponding to or its complement at positions 440-446 as described in SEQ ID NO: 16, the position corresponding to or its complement at positions 361-367 as described in SEQ ID NO: 21, the position corresponding to or its complement at positions 600-606 as described in SEQ ID NO: 26, the position corresponding to or its complement at positions 539-545 as described in SEQ ID NO: 35, the position corresponding to or its complement at positions 440-446 as described in SEQ ID NO: 40, the position corresponding to or its complement at positions 361-367 as described in SEQ ID NO: 45, or the position corresponding to or its complement at positions 600-606 as described in SEQ ID NO: 50.

[0180] In some embodiments, the isolated change-specific probe or change-specific primer comprises at least about 15 nucleotides, the change-specific probe or change-specific primer comprising a nucleotide sequence complementary to a portion of the nucleotide sequence encoding the human GPR75 polypeptide, the portion comprising the position corresponding to or its complement at position 5,557 as described in SEQ ID NO: 3, the position corresponding to or its complement at position 556 as described in SEQ ID NO: 12, the position corresponding to or its complement at position 457 as described in SEQ ID NO: 17, the position corresponding to or its complement at position 378 as described in SEQ ID NO: 22, the position corresponding to or its complement at position 617 as described in SEQ ID NO: 27, the position corresponding to or its complement at position 556 as described in SEQ ID NO: 36, the position corresponding to or its complement at position 457 as described in SEQ ID NO: 41, the position corresponding to or its complement at position 378 as described in SEQ ID NO: 46, or the position corre...

Claims

1. An in vitro method for testing the susceptibility of a subject to obesity, This includes determining, or having determined, the presence or absence of a G protein-coupled receptor 75 (GPR75) missense variant nucleic acid molecule encoding a predicted loss-of-function GPR75 polypeptide in a biological sample obtained from the subject, The fact that the subject is GPR75 referenced is an indicator that the subject is at high risk of developing obesity. The in vitro method wherein the subject being heterozygous or homozygous for a GPR75 missense variant nucleic acid molecule is an indicator that the subject has a low risk of developing obesity.

2. The in vitro method according to claim 1, wherein the GPR75 missense variant nucleic acid molecule comprises a nucleic acid molecule encoding Ala110fs, Ala116Thr, Tyr207Cys, Glun234Stop, Arg236fs, or Cys400fs.

3. The aforementioned GPR75 missense variant nucleic acid molecule, It lacks the CCAGTAG heptanucleotide at the position corresponding to positions 5,540-5,546 as described in Sequence ID No.

1. The adenine at the position corresponding to position 5,557 as described in Sequence ID No. 3, Including the thymine at the position corresponding to position 5,911 as described in Sequence ID No. 4, It lacks the AAAG tetranucleotide at the position corresponding to positions 5,920-5,923 as described in Sequence ID No.

1. This includes the insertion of a thymine at the position corresponding to position 6,411 as described in Sequence ID No. 6, or Contains guanine at the position corresponding to position 5,831 as described in Sequence ID No. 99, A genome nucleic acid molecule having a nucleotide sequence, It lacks the CCAG heptanucleotide at positions corresponding to 539-545 as described in Sequence ID No.

7. It lacks the CCAG heptanucleotide at positions corresponding to positions 440-446 as described in Sequence ID No.

8. It lacks the CCAG heptanucleotide at positions corresponding to positions 361-367 as described in Sequence ID No.

9. It lacks the CCAG heptanucleotide at the position corresponding to positions 600-606 as described in Sequence ID No.

10. The adenine at the position corresponding to position 556 as described in Sequence ID No. 12, Contains adenine at the position corresponding to position 457 as described in Sequence ID No. 17, The adenine at position 378, as described in Sequence ID No. 22, The adenine at the position corresponding to position 617 described in Sequence ID No. 27, Contains uracil at the position corresponding to position 910 as described in Sequence ID No. 13, Includes uracil at the position corresponding to position 811 as described in Sequence ID No. 18, Includes uracil at position 732, as described in Sequence ID No. 23, Includes uracil at position 971, as described in Sequence ID No. 28, It lacks the AAAG tetranucleotide at the position corresponding to positions 919-922 as described in Sequence ID No.

7. It lacks the AAAG tetranucleotide at the position corresponding to positions 820-823 as described in Sequence ID No.

8. It lacks the AAAG tetranucleotide at positions corresponding to positions 741-744 as described in Sequence ID No.

9. It lacks the AAAG tetranucleotide at the position corresponding to positions 980-983 as described in Sequence ID No.

10. Including the insertion of uracil at the position corresponding to position 1,410 as described in Sequence ID No. 15, Including the insertion of uracil at the position corresponding to position 1,311 as described in Sequence ID No. 20, Including the insertion of uracil at the position corresponding to position 1,232 as described in Sequence ID No. 25, Including the insertion of uracil at the position corresponding to position 1,471 as described in Sequence ID No. 30, The guanine at the position corresponding to position 830 described in Sequence ID No. 100 is Containing guanine at the position corresponding to position 731 as described in Sequence ID No. 101, Contains guanine at the position corresponding to position 652 as described in Sequence ID No. 102, or Containing guanine at the position corresponding to position 891 as described in Sequence ID No. 103, mRNA molecules having a nucleotide sequence, or cDNA molecules generated from mRNA molecules, It lacks the CCAGTAG heptanucleotide at positions corresponding to 539-545 as described in Sequence ID No.

31. It lacks the CCAGTAG heptanucleotide at positions corresponding to 440-446 as described in Sequence ID No.

32. It lacks the CCAGTAG heptanucleotide at positions corresponding to positions 361-367 as described in SEQ ID NO:

33. It lacks the CCAGTAG heptanucleotide at the position corresponding to positions 600-606 as described in Sequence ID No.

34. The adenine at the position corresponding to position 556 described in Sequence ID No. 36, The adenine at the position corresponding to position 457 described in Sequence ID No. 41, Contains adenine at the position corresponding to position 378 as described in Sequence ID No. 46, The adenine at the position corresponding to position 617 described in Sequence ID No. 51, Including the thymine at the position corresponding to position 910 as described in Sequence ID No. 37, Including the thymine at the position corresponding to position 811 as described in Sequence ID No. 42, Including the thymine at position 732, as described in Sequence ID No. 47, Including the thymine at position 971, as described in Sequence ID No. 52, It lacks the AAAG tetranucleotide at the position corresponding to positions 919-922 as described in Sequence ID No.

31. It lacks the AAAG tetranucleotide at the position corresponding to positions 820-823 as described in Sequence ID No.

32. It lacks the AAAG tetranucleotide at positions corresponding to positions 741-744 as described in Sequence ID No.

33. It lacks the AAAG tetranucleotide at the position corresponding to positions 980-983 as described in Sequence ID No.

34. Including the insertion of thymine at the position corresponding to position 1,410 as described in Sequence ID No. 39, Including the insertion of thymine at the position corresponding to position 1,311 as described in Sequence ID No. 44, Including the insertion of thymine at the position corresponding to position 1,232 as described in Sequence ID No. 49, Including the insertion of thymine at the position corresponding to position 1,471 as described in Sequence ID No. 54, Containing guanine at the position corresponding to position 830 as described in Sequence ID No. 104, Containing guanine at the position corresponding to position 731 as described in Sequence ID No. 105, Contains guanine at the position corresponding to position 652 as described in Sequence ID No. 106, or Containing guanine at the position corresponding to position 891 as described in Sequence ID No. 107, The cDNA molecule having a nucleotide sequence The in vitro method according to claim 2, including the following:

4. A genomic nucleic acid molecule having a nucleotide sequence encoding a human G protein-coupled receptor 75 (GPR75) polypeptide, wherein the nucleotide sequence is It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in Sequence ID No.

1. The adenine or its complement at the position corresponding to position 5,557 as described in Sequence ID No. 3, The thymine or its complement at the position corresponding to position 5,911 as described in Sequence ID No. 4, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 5,920-5,923 as described in Sequence ID No. 1, This includes the insertion of thymine or its complement at the position corresponding to position 6,411 as described in Sequence ID No. 6, or The compound includes guanine or its complement at the position corresponding to position 5,831 as described in Sequence ID No.

99. The aforementioned genome nucleic acid molecule, An mRNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence is It lacks the CCAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in Sequence ID No.

7. Lack of the CCAG heptanucleotide or its complement at positions 440-446, as described in Sequence ID No.

8. Lack of CCAG heptanucleotide or its complement at positions 361-367, as described in Sequence ID No.

9. Lack of the CCAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in Sequence ID No. 10, The adenine or its complement at the position corresponding to position 556 as described in Sequence ID No. 12, The adenine or its complement at the position corresponding to position 457 as described in Sequence ID No. 17, The adenine or its complement at the position corresponding to position 378 as described in Sequence ID No. 22, The adenine or its complement at the position corresponding to position 617 as described in Sequence ID No. 27, Includes uracil or its complement at the position corresponding to position 910 as described in Sequence ID No. 13, Includes uracil or its complement at the position corresponding to position 811 as described in Sequence ID No. 18, Includes uracil or its complement at the position corresponding to position 732 as described in Sequence ID No. 23, Includes uracil or its complement at the position corresponding to position 971 as described in Sequence ID No. 28, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in Sequence ID No. 7, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in Sequence ID No. 8, Lacking the AAAG tetranucleotide or its complement at positions corresponding to positions 741-744 as described in Sequence ID No. 9, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in Sequence ID No. 10, Includes insertion of uracil or its complement at the position corresponding to position 1,410 as described in Sequence ID No. 15, Includes insertion of uracil or its complement at the position corresponding to position 1,311 as described in Sequence ID No. 20, Includes insertion of uracil or its complement at the position corresponding to position 1,232 as described in Sequence ID No. 25, Includes insertion of uracil or its complement at the position corresponding to position 1,471 as described in Sequence ID No. 30, Includes guanine or its complement at the position corresponding to position 830 as described in Sequence ID No. 100, Includes guanine or its complement at the position corresponding to position 731 as described in Sequence ID No. 101, It contains guanine or its complement at the position corresponding to position 652 as described in Sequence ID No. 102, or Includes guanine or its complement at the position corresponding to position 891 as described in Sequence ID No. 103, The mRNA molecule, or A cDNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence is It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in Sequence ID No.

31. It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in Sequence ID No.

32. It lacks the CCAGTAG heptanucleotide or its complement at the positions corresponding to positions 361-367 as described in Sequence ID No.

33. Lack of the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in Sequence ID No. 34, The adenine or its complement at the position corresponding to position 556 as described in Sequence ID No. 36, The adenine or its complement at the position corresponding to position 457 as described in Sequence ID No. 41, The adenine or its complement at the position corresponding to position 378 as described in Sequence ID No. 46, The adenine or its complement at the position corresponding to position 617 as described in Sequence ID No. 51, The thymine or its complement at the position corresponding to position 910 as described in Sequence ID No. 37, The thymine or its complement at the position corresponding to position 811 as described in Sequence ID No. 42, The thymine or its complement at the position corresponding to position 732 as described in Sequence ID No. 47, The thymine or its complement at the position corresponding to position 971 as described in Sequence ID No. 52, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in Sequence ID No. 31, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in Sequence ID No. 32, Lacking the AAAG tetranucleotide or its complement at positions corresponding to positions 741-744 as described in Sequence ID No. 33, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in Sequence ID No. 34, Includes the insertion of thymine or its complement at the position corresponding to position 1,410 as described in Sequence ID No. 39, Includes the insertion of thymine or its complement at the position corresponding to position 1,311 as described in Sequence ID No. 44, Includes the insertion of thymine or its complement at the position corresponding to position 1,232 as described in Sequence ID No. 49, Includes the insertion of thymine or its complement at the position corresponding to position 1,471 as described in Sequence ID No. 54, Includes guanine or its complement at the position corresponding to position 830 as described in Sequence ID No. 104, The guanine or its complement at the position corresponding to position 731 as described in Sequence ID No. 105, It contains guanine or its complement at the position corresponding to position 652 as described in Sequence ID No. 106, or Includes guanine or its complement at the position corresponding to position 891 as described in Sequence ID No. 107, The cDNA molecule The use of a G protein-coupled receptor 75 (GPR75) inhibitor in the manufacture of a pharmaceutical for treating obesity in a subject determined to have the following, wherein the GPR75 inhibitor comprises an inhibitory nucleic acid molecule that hybridizes with GPR75 mRNA.

5. The use according to claim 4, wherein the inhibitory nucleic acid molecule includes an antisense nucleic acid molecule that hybridizes with GPR75 mRNA, a small interfering RNA (siRNA) that hybridizes with GPR75 mRNA, or a short hairpin RNA (shRNA) that hybridizes with GPR75 mRNA.

6. A genomic nucleic acid molecule having a nucleotide sequence encoding a human G protein-coupled receptor 75 (GPR75) polypeptide, wherein the nucleotide sequence is It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 5,540-5,546 as described in Sequence ID No.

1. The adenine or its complement at the position corresponding to position 5,557 as described in Sequence ID No. 3, The thymine or its complement at the position corresponding to position 5,911 as described in Sequence ID No. 4, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 5,920-5,923 as described in Sequence ID No. 1, This includes the insertion of thymine or its complement at the position corresponding to position 6,411 as described in Sequence ID No. 6, or The compound includes guanine or its complement at the position corresponding to position 5,831 as described in Sequence ID No.

99. The aforementioned genome nucleic acid molecule, An mRNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence is It lacks the CCAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in Sequence ID No.

7. Lack of the CCAG heptanucleotide or its complement at positions 440-446, as described in Sequence ID No.

8. Lack of CCAG heptanucleotide or its complement at positions 361-367, as described in Sequence ID No.

9. Lack of the CCAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in Sequence ID No. 10, The adenine or its complement at the position corresponding to position 556 as described in Sequence ID No. 12, The adenine or its complement at the position corresponding to position 457 as described in Sequence ID No. 17, The adenine or its complement at the position corresponding to position 378 as described in Sequence ID No. 22, The adenine or its complement at the position corresponding to position 617 as described in Sequence ID No. 27, Includes uracil or its complement at the position corresponding to position 910 as described in Sequence ID No. 13, Includes uracil or its complement at the position corresponding to position 811 as described in Sequence ID No. 18, Includes uracil or its complement at the position corresponding to position 732 as described in Sequence ID No. 23, Includes uracil or its complement at the position corresponding to position 971 as described in Sequence ID No. 28, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in Sequence ID No. 7, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 820-823 as described in Sequence ID No. 8, Lacking the AAAG tetranucleotide or its complement at positions corresponding to positions 741-744 as described in Sequence ID No. 9, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in Sequence ID No. 10, Includes insertion of uracil or its complement at the position corresponding to position 1,410 as described in Sequence ID No. 15, Includes insertion of uracil or its complement at the position corresponding to position 1,311 as described in Sequence ID No. 20, Includes insertion of uracil or its complement at the position corresponding to position 1,232 as described in Sequence ID No. 25, Includes insertion of uracil or its complement at the position corresponding to position 1,471 as described in Sequence ID No. 30, Includes guanine or its complement at the position corresponding to position 830 as described in Sequence ID No. 100, Includes guanine or its complement at the position corresponding to position 731 as described in Sequence ID No. 101, It contains guanine or its complement at the position corresponding to position 652 as described in Sequence ID No. 102, or Includes guanine or its complement at the position corresponding to position 891 as described in Sequence ID No. 103, The mRNA molecule, or A cDNA molecule having a nucleotide sequence encoding a human GPR75 polypeptide, wherein the nucleotide sequence is It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 539-545 as described in Sequence ID No.

31. It lacks the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 440-446 as described in Sequence ID No.

32. It lacks the CCAGTAG heptanucleotide or its complement at the positions corresponding to positions 361-367 as described in Sequence ID No.

33. Lack of the CCAGTAG heptanucleotide or its complement at the position corresponding to positions 600-606 as described in Sequence ID No. 34, The adenine or its complement at the position corresponding to position 556 as described in Sequence ID No. 36, The adenine or its complement at the position corresponding to position 457 as described in Sequence ID No. 41, The adenine or its complement at the position corresponding to position 378 as described in Sequence ID No. 46, The adenine or its complement at the position corresponding to position 617 as described in Sequence ID No. 51, The thymine or its complement at the position corresponding to position 910 as described in Sequence ID No. 37, The thymine or its complement at the position corresponding to position 811 as described in Sequence ID No. 42, The thymine or its complement at the position corresponding to position 732 as described in Sequence ID No. 47, The thymine or its complement at the position corresponding to position 971 as described in Sequence ID No. 52, Lacking the AAAG tetranucleotide or its complement at the positions corresponding to positions 919-922 as described in Sequence ID No. 31, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 820-823 as described in Sequence ID No. 32, Lacking the AAAG tetranucleotide or its complement at positions corresponding to positions 741-744 as described in Sequence ID No. 33, Lacking the AAAG tetranucleotide or its complement at the position corresponding to positions 980-983 as described in Sequence ID No. 34, Includes the insertion of thymine or its complement at the position corresponding to position 1,410 as described in Sequence ID No. 39, Includes the insertion of thymine or its complement at the position corresponding to position 1,311 as described in Sequence ID No. 44, Includes the insertion of thymine or its complement at the position corresponding to position 1,232 as described in Sequence ID No. 49, Includes the insertion of thymine or its complement at the position corresponding to position 1,471 as described in Sequence ID No. 54, Includes guanine or its complement at the position corresponding to position 830 as described in Sequence ID No. 104, The guanine or its complement at the position corresponding to position 731 as described in Sequence ID No. 105, It contains guanine or its complement at the position corresponding to position 652 as described in Sequence ID No. 106, or Includes guanine or its complement at the position corresponding to position 891 as described in Sequence ID No. 107, The cDNA molecule The use of a G protein-coupled receptor 75 (GPR75) inhibitor in the manufacture of a pharmaceutical product for treating obesity in a subject determined to have the following, wherein the GPR75 inhibitor comprises a Cas protein and a gRNA that hybridizes with a guide RNA (gRNA) recognition sequence within the GPR75 genomic nucleic acid molecule.

7. The use according to claim 6, wherein the Cas protein comprises Cas9 or Cpf1.

8. The use according to claim 6 or 7, wherein the gRNA recognition sequence includes or is adjacent to a position corresponding to position 5,540-5,546, position 5,557, position 5,911, position 5,920-5,923, position 6,411, or position 5,831 as described in SEQ ID NO:

1.

9. The use according to any one of claims 6 to 8, wherein the gRNA comprises about 17 to about 23 nucleotides.

10. The use according to any one of claims 6 to 9, wherein the gRNA recognition sequence includes the nucleotide sequence described in any one of sequence numbers 61 to 98.

11. The in vitro method according to any one of claims 1 to 3, wherein the subject is a human.

12. The in vitro method according to claim 1, wherein the GPR75 missense variant nucleic acid molecule does not encode GPR75 Ala116Thr.

13. Use of a G protein-coupled receptor 75 (GPR75) inhibitor in the manufacture of a pharmaceutical product for treating obesity in a subject determined to be GPR75-referenced, wherein the GPR75 inhibitor comprises an inhibitory nucleic acid molecule that hybridizes with GPR75 mRNA.

14. The use according to any one of claims 4 to 10 and 13, wherein the subject is a human.

15. Use of a G protein-coupled receptor 75 (GPR75) inhibitor in the manufacture of a pharmaceutical product for treating obesity of a subject determined to be GPR75-referenced, wherein the GPR75 inhibitor comprises a Cas protein and a gRNA that hybridizes with a guide RNA (gRNA) recognition sequence within the GPR75 genomic nucleic acid molecule.

16. The use according to claim 15, wherein the Cas protein comprises Cas9 or Cpf1.

17. The use according to claim 15 or claim 16, wherein the gRNA recognition sequence includes or is adjacent to a position corresponding to position 5,540-5,546, position 5,557, position 5,911, position 5,920-5,923, position 6,411, or position 5,831 as described in SEQ ID NO:

1.

18. The use according to any one of claims 15 to 17, wherein the gRNA comprises about 17 to about 23 nucleotides.

19. The use according to any one of claims 15 to 18, wherein the gRNA recognition sequence includes the nucleotide sequence described in any one of sequence numbers 61 to 98.

20. The use according to any one of claims 15 to 19, wherein the subject is a human.

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