Polynucleotides encoding novel variants of the TRP channel family member, LTRPC3

• US 20040229315A1
• Filed: 05/10/2004
• Published: 11/18/2004
• Est. Priority Date: 05/12/2003
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Chen, Jian, Feder, John N., Wu, Shujian, Blanar, Michael A., Sun, Lucy, Lee, Ning, Levesque, Paul C., Bol, David K.
• Granted Patent: US 7,344,882 B2
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)

Abstract

The present invention provides novel polynucleotides encoding LTRPC3g, LTRPC3h, LTRPC3i, LTRPC3j, LTRPC3k, or LTRPC3l polypeptides, fragments and homologues thereof. Also provided are vectors, host cells, antibodies, and recombinant and synthetic methods for producing said polypeptides. The invention further relates to diagnostic and therapeutic methods for applying these novel LTRPC3g, LTRPC3h, LTRPC3i, LTRPC3j, LTRPC3k, or LTRPC3l polypeptides to the diagnosis, treatment, and/or prevention of various diseases and/or disorders related to these polypeptides. The invention further relates to screening methods for identifying agonists and antagonists of the polynucleotides and polypeptides of the present invention.

Claims

1. An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence selected from the group consisting of:

(a) an isolated polynucleotide comprising nucleotides 75 to 5198 of SEQ ID NO;

1, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1709 of SEQ ID NO;

2;

(b) an isolated polynucleotide comprising nucleotides 72 to 5198 of SEQ ID NO;

1, wherein said nucleotides encode a polypeptide corresponding to amino acids 1 to 1709 of SEQ ID NO;

2;

(c) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

1;

(d) an isolated polynucleotide comprising nucleotides 75 to 5234 of SEQ ID NO;

3, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1721 of SEQ ID NO;

4;

(e) an isolated polynucleotide comprising nucleotides 72 to 5234 of SEQ ID NO;

3, wherein said nucleotides encode a polypeptide corresponding to amino acids 1 to 1721 of SEQ ID NO;

4;

(f) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

3;

(g) an isolated polynucleotide comprising nucleotides 75 to 5234 of SEQ ID NO;

5, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1721 of SEQ ID NO;

6;

(h) an isolated polynucleotide comprising nucleotides 72 to 5234 of SEQ ID NO;

5, wherein said nucleotides encode a polypeptide corresponding to amino acids 1 to 1721 of SEQ ID NO;

6;

(i) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

5;

(j) an isolated polynucleotide comprising nucleotides 75 to 5168 of SEQ ID NO;

7, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1699 of SEQ ID NO;

8;

(k) an isolated polynucleotide comprising nucleotides 72 to 5168 of SEQ ID NO;

7, wherein said nucleotides encode a polypeptide corresponding to amino acids 1 to 1699 of SEQ ID NO;

8;

(l) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

7;

(m) an isolated polynucleotide comprising nucleotides 75 to 5204 of SEQ ID NO;

9, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1711 of SEQ ID NO;

10;

(n) an isolated polynucleotide comprising nucleotides 72 to 5204 of SEQ ID NO;

9, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1711 of SEQ ID NO;

10;

(o) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

9;

(p) an isolated polynucleotide comprising nucleotides 75 to 5273 of SEQ ID NO;

11, wherein said nucleotides encode a polypeptide corresponding to amino acids 2 to 1734 of SEQ ID NO;

12;

(q) an isolated polynucleotide comprising nucleotides 72 to 5273 of SEQ ID NO;

11, wherein said nucleotides encode a polypeptide corresponding to amino acids 1 to 1734 of SEQ ID NO;

12;

(r) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

11; and

(s) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(r), wherein said polynucleotide does not hybridize under stringent conditions to a nucleic acid molecule having a nucleotide sequence of only A residues or of only T residues.

2. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide comprises a nucleotide sequence encoding a human transient receptor potential channel.

3. A recombinant vector comprising the isolated nucleic acid molecule of claim 1.

4. A recombinant host cell comprising the vector sequences of claim 3.

12. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:

(a) determining the presence or absence of a mutation in the polynucleotide of claim 1;

and

(b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or absence of said mutation.

5. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:

(a) a polypeptide comprising amino acids 2 to 1709 of SEQ ID NO;

2;

(b) a polypeptide comprising amino acids 1 to 1709 of SEQ ID NO;

2;

(c) a polypeptide comprising amino acids 2 to 1721 of SEQ ID NO;

4;

(d) a polypeptide comprising amino acids 1 to 1721 of SEQ ID NO;

4;

(e) a polypeptide comprising amino acids 2 to 1721 of SEQ ID NO;

6;

(f) a polypeptide comprising amino acids 1 to 1721 of SEQ ID NO;

6;

(g) a polypeptide comprising amino acids 2 to 1699 of SEQ ID NO;

8;

(h) a polypeptide comprising amino acids 1 to 1699 of SEQ ID NO;

8;

(i) a polypeptide comprising amino acids 2 to 1711 of SEQ ID NO;

10;

(j) a polypeptide comprising amino acids 1 to 1711 of SEQ ID NO;

10;

(k) a polypeptide comprising amino acids 2 to 1734 of SEQ ID NO;

12; and

(l) a polypeptide comprising amino acids 1 to 1734 of SEQ ID NO;

12.

6. The isolated polypeptide of claim 5, wherein the full length protein comprises sequential amino acid deletions from either the C-terminus or the N-terminus.

7. An isolated antibody that binds specifically to the isolated polypeptide of claim 5.

8. A recombinant host cell that expresses the isolated polypeptide of claim 5.

9. A method of making an isolated polypeptide comprising:

(a) culturing the recombinant host cell of claim 8 under conditions such that said polypeptide is expressed; and

(b) recovering said polypeptide.

10. The polypeptide produced by claim 9.

11. A method for preventing, treating, or ameliorating a medical condition, comprising the step of administering to a mammalian subject a therapeutically effective amount of the polypeptide of claim 5, or a modulator thereof.

13. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:

(a) determining the presence or amount of expression of the polypeptide of claim 5 in a sample; and

(b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or amount of expression of the polypeptide.

19. The method for preventing, treating, or ameliorating a medical condition of claim 11, wherein the medical condition is selected from the group consisting of a renal disorder;

a disorder related to aberrant calcium regulation;

a reproductive disorder;

a neural disorder;

cerebellum disorders;

various choroid plexus neoplasms;

choroid plexus papilloma;

Alzheimer'"'"'s disease;

prion disorders;

multiple sclerosis;

movement disorders that involve the cerebellum;

a proliferative kidney or renal disorder;

solitary metastasis of the kidney;

renal cell carcinoma;

ovarian cancer or related proliferative condition of the ovary;

testicular cancer or related proliferative condition of the testis;

kidney cancer or related proliferative condition of the kidney;

a disorder that maps to or is associated with chromosome locus 9q21.11-21.31;

amyotrophic lateral sclerosis with frontotemporal dementia;

early-onset pulverulent cataract;

infantile nephronophthisis;

hypomagnesemia with secondary hypocalcemia and/or familial hemophagocytic lymphohistiocytosis;

a disorder associated with disturbances in Ca²⁺ homeostasis including osteoporosis;

hypercalciuric stone disease;

chronic renal failure;

a condition that would benefit from modulation of intracellular Ca++ ion concentrations;

a condition that would benefit from modulation of Ca++ ion flux;

a condition that would benefit from modulation of stored intracellular Ca++ ion concentrations;

a condition that would benefit from modulation of Ca++ ion pump activity;

a condition that would benefit from modulation of Ca++ ion flow into a cell;

a condition that would benefit from modulation of Ca++ ion flow out of cells;

a condition that would benefit from activation of Ca++ senstive proteins;

a condition that would benefit from the activation of Ca++ senstive signaling pathways;

the activation of kinase-activatible proteins;

a condition that would benefit from the activation of kinase-dependent signaling pathways;

DNA-repair deficiencies, particularly base-excision repair deficiencies;

Xeroderma pigmentosum;

skin cancer;

melanoma;

UV senstivity;

alkylation sensivity;

gamma irradiation sensitivity;

pyrimidine dimer sensitivity;

chemical mutagenesis;

lymphomas, leukemias;

photosensitivity;

Bloom'"'"'s syndrone;

Fanconi'"'"'s anemia;

ataxia telangiectasia;

chromosomal aberrations;

blood vessel dilation aberrations in the skin;

blood vessel dilation aberrations in the eye;

conditions involving increased levels of apurinic sites;

conditions involving increased levels of apyrimidinic sites;

conditions involving increased levels of abasic sites;

disorders related to aberrant signal transduction;

proliferating disorders;

cancers;

a disorder related to misregulation of FEN1expression or activity;

a disorder that would benefit from increasing mammalian base excision repair activity;

increasing mammalian single-nucleotide base excision repair activity; and

increasing mammalian long patch base excision repair activity.

20. The method for preventing, treating, or ameliorating a medical condition of claim 11, wherein the medical condition is selected from the group consisting of:

Ca²⁺ reabsorption disorders, disorders associated with aberrant luminal Ca2+ entry via the epithelial Ca2+ channel (ECaC), disorders associated with aberrant cytosolic diffusion of Ca2+ bound to calbindin-D28K, disorders associated with aberrant basolateral extrusion of Ca2+ through the Na+/Ca2+ exchanger (NCX), disorders associated with aberrant plasma membrane Ca2+-ATPase (PMCA) activity and/or activation, disorders associated with the hypocalciuric effect of diuretics, disorders associated with the hypocalciuric effect of thiazide diuretics, disorders associated with hormone insufficiencies that affect the function of the kidney, disorders associated with hormone insufficiencies that affect renal Ca2+ homeostasis, disorders associated with aberrant renal Ca2+ homeostasis as a result of vitamin D deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of PTH deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of vasopressin deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of prostaglandines deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of estrogen deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of calcitonin deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of parathyroid hormone deficiency, disorders associated with aberrant renal Ca2+ homeostasis as a result of atrial natriuretic peptide deficiency, calcium homeostasis-related disorders, hypercaliuric nephrolithiasis, certain forms of osteoporosis, Gitelman'"'"'s disease, Bartter'"'"'s syndrom, disorders associated with aberrant function of kidney tubular epithelium, disorders associated with aberrant function of kidney tubular epithelium in the medulla, disorders associated with aberrant function of kidney tubular epithelium in the medullary rays, disorders associated with aberrant function of kidney tubular epithelium in the periglomerular, acute kidney failure, chronic kidney failure, cystic kidney disease, horseshoe kidney, hypertensive kidney disease, kidney atrophy, kidney cancer, kidney disease, kidney failure, kidney infection, kidney metastasis, kidney stone, nephrolithiasis, multicystic dysplastic kidney disease, and polycystic kidney disease.

14. An isolated nucleic acid molecule consisting of a polynucleotide having a nucleotide sequence selected from the group consisting of:

(a) an isolated polynucleotide consisting of nucleotides 75 to 5198 of SEQ ID NO;

1;

(b) an isolated polynucleotide consisting of nucleotides 72 to 5198 of SEQ ID NO;

1;

(c) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

1;

(d) an isolated polynucleotide consisting of nucleotides 75 to 5234 of SEQ ID NO;

3;

(e) an isolated polynucleotide consisting of nucleotides 72 to 5234 of SEQ ID NO;

3;

(f) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

3;

(g) an isolated polynucleotide consisting of nucleotides 75 to 5234 of SEQ ID NO;

5;

(h) an isolated polynucleotide consisting of nucleotides 72 to 5234 of SEQ ID NO;

5;

(i) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

5;

(j) an isolated polynucleotide consisting of nucleotides 75 to 5168 of SEQ ID NO;

7;

(k) an isolated polynucleotide consisting of nucleotides 72 to 5168 of SEQ ID NO;

7;

(l) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

7;

(m) an isolated polynucleotide consisting of nucleotides 75 to 5204 of SEQ ID NO;

9;

(n) an isolated polynucleotide consisting of nucleotides 72 to 5204 of SEQ ID NO;

9;

(o) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

9;

(p) an isolated polynucleotide consisting of nucleotides 75 to 5273 of SEQ ID NO;

11;

(q) an isolated polynucleotide consisting of nucleotides 72 to 5273 of SEQ ID NO;

11; and

(r) a polynucleotide which represents the complimentary sequence of SEQ ID NO;

11.

15. The isolated nucleic acid molecule of claim 14, wherein the polynucleotide comprises a nucleotide sequence encoding a human transient receptor potential channel.

16. A recombinant vector comprising the isolated nucleic acid molecule of claim 15.

17. A recombinant host cell comprising the recombinant vector of claim 16.

18. An isolated polypeptide consisting of an amino acid sequence selected from the group consisting of:

(a) a polypeptide corresponding to amino acids 2 to 1709 of SEQ ID NO;

2;

(b) a polypeptide corresponding to amino acids 1 to 1709 of SEQ ID NO;

2;

(c) a polypeptide corresponding to amino acids 2 to 1721 of SEQ ID NO;

4;

(d) a polypeptide corresponding to amino acids 1 to 1721 of SEQ ID NO;

4;

(e) a polypeptide corresponding to amino acids 2 to 1721 of SEQ ID NO;

6;

(f) a polypeptide corresponding to amino acids 1 to 1721 of SEQ ID NO;

6;

(g) a polypeptide corresponding to amino acids 2 to 1699 of SEQ ID NO;

8;

(h) a polypeptide corresponding to amino acids 1 to 1699 of SEQ ID NO;

8;

(i) a polypeptide corresponding to amino acids 2 to 1711 of SEQ ID NO;

10;

(j) a polypeptide corresponding to amino acids 1 to 1711 of SEQ ID NO;

10;

(k) a polypeptide corresponding to amino acids 2 to 1734 of SEQ ID NO;

12; and

(l) a polypeptide corresponding to amino acids 1 to 1734 of SEQ ID NO;

12.