Human ion channels
OwnerSteven L. Roberds, Christopher W. Benjamin +5inventors
InventorsSteven L. Roberds, Christopher W. Benjamin, Alla M. Karnovsky, Cara L. Ruble, Lisa I. Linske-O'Connell, Jun Wang, Derong Liu
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Abstract
The present invention provides novel ion channel polypeptides and polynucleotides which identify and encode them. In addition, the invention provides expression vectors, host cells and methods for their production. The invention also provides methods for the identification of ion channel agonists/antagonists, useful for the treatment of human diseases and conditions.
Claims116
An isolated nucleic acid molecule comprising a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence homologous to a sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110, said nucleic acid molecule encoding at least a portion of ion-x.
An isolated nucleic acid molecule comprising at least 10 nucleotides, said nucleic acid molecule comprising a nucleotide sequence complementary to a sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:51, and SEQ ID NOS:107 and 108.
A method for identifying a compound which binds ion-x comprising the steps of:
- a) contacting ion-x with a compound;
- and
- b) determining whether said compound binds ion-x.
A method for identifying a compound which binds a nucleic acid molecule encoding ion-x comprising the steps of:
- a) contacting said nucleic acid molecule encoding ion-x with a compound;
- and
- b) determining whether said compound binds said nucleic acid molecule.
A method for identifying a compound which modulates the activity of ion-x comprising the steps of:
- a) contacting ion-x with a compound;
- and
- b) determining whether ion-x activity has been modulated.
A method of identifying an animal homolog of ion-x comprising the steps:
- a) comparing the nucleic acid sequences of the animal with a sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:51, and SEQ ID NOS:107 and 108;
- and
- b) identifying nucleic acid sequences of the animal that are homologous to said sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:51, and SEQ ID NOS:107 and 108.
A method of screening a human subject to diagnose a disorder affecting the brain or genetic predisposition therefor, comprising the steps of:
- (a) assaying nucleic acid of a human subject to determine a presence or an absence of a mutation altering an amino acid sequence, expression, or biological activity of at least one ion channel that is expressed in the brain, wherein the ion channel comprises an amino acid sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110, and allelic variants thereof, and wherein the nucleic acid corresponds to a gene encoding the ion channel;
- and
- (b) diagnosing the disorder or predisposition from the presence or absence of said mutation, wherein the presence of a mutation altering the amino acid sequence, expression, or biological activity of the ion channel correlates with an increased risk of developing the disorder.
A method of screening for an ion-x mental disorder genotype in a human patient, comprising the steps of:
- (a) providing a biological sample comprising nucleic acid from said patient, said nucleic acid including sequences corresponding to alleles of ion-x;
- and
- (b) detecting the presence of one or more mutations in the ion-x alleles;
- wherein the presence of a mutation in an ion-x allele is indicative of a mental disorder genotype.
A kit for screening a human subject to diagnose a mental disorder or a genetic predisposition therefor, comprising, in association:
- (a) an oligonucleotide useful as a probe for identifying polymorphisms in a human ion-x gene, the oligonucleotide comprising 6-50 nucleotides in a sequence that is identical or complementary to a sequence of a wild type human ion-x coding sequence, except for one sequence difference selected from the group consisting of a nucleotide addition, a nucleotide deletion, or nucleotide substitution;
- and
- (b) a media packaged with the oligonucleotide, said media containing information for identifying polymorphisms that correlate with a mental disorder or a genetic predisposition therefor, the polymorphisms being identifiable using the oligonucleotide as a probe.
A method of identifying an ion channel allelic variant that correlates with a mental disorder, comprising steps of:
- (a) providing a biological sample comprising nucleic acid from a human patient diagnosed with a mental disorder, or from the patient's genetic progenitors or progeny;
- (b) detecting in the nucleic acid the presence of one or more mutations in an ion channel that is expressed in the brain, wherein the ion channel comprises an amino acid sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110, and allelic variants thereof, and wherein the nucleic acid includes sequence corresponding to the gene or genes encoding the ion channel;
- wherein the one or more mutations detected indicates an allelic variant that correlates with a mental disorder.
A purified polynucleotide comprising a nucleotide sequence encoding ion-x of a human with a mental disorder;
- wherein said polynucleotide hybridizes to the complement of a sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110 under the following hybridization conditions:
- (a) hybridization for 16 hours at 42° C. in a hybridization solution comprising 50% formamide, 1% SDS, 1 M NaCl, 10% dextran sulfate and
- (b) washing 2 times for 30 minutes at 60° C. in a wash solution comprising 0.1×SSC and 1% SDS;
- and
- wherein the polynucleotide that encodes ion-x amino acid sequence of the human differs from the sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110 by at least one residue.
A method to identify compounds useful for the treatment of a disorder, said method comprising the steps of:
- (a) contacting a composition comprising ion-x with a compound suspected of binding ion-x;
- (b) detecting binding between ion-x and the compound suspected of binding ion-x;
- wherein compounds identified as binding ion-x are candidate compounds useful for the treatment of a disorder.
A method for identifying a compound useful as a modulator of binding between ion-x and a binding partner of ion-x comprising the steps of:
- (a) contacting the binding partner and a composition comprising ion-x in the presence and in the absence of a putative modulator compound;
- (b) detecting binding between the binding partner and ion-x;
- wherein decreased or increased binding between the binding partner and ion-x in the presence of the putative modulator, as compared to binding in the absence of the putative modulator is indicative a modulator compound useful for the treatment of a disorder.
A chimeric receptor comprising at least 5 amino acid residues, said receptor comprising at least a portion of a sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102, and SEQ ID NOS:109 and 110.
An isolated nucleic acid molecule comprising a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence homologous to a sequence selected from the group consisting of SEQ ID NO:52 to SEQ ID NO:102 and SEQ ID NOS:109 and 110, said nucleic acid molecule encoding at least a portion of ion-x.
An isolated nucleic acid molecule comprising a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence homologous to a sequence selected from the group consisting of SEQ ID NO:105 and SEQ ID NO:106.
An isolated nucleic acid molecule comprising a nucleotide sequence homologous to a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104.
78 An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104.
83 A host cell transformed with an expression vector of claim 81.
84 An isolated nucleic acid molecule comprising at least 10 nucleotides, said nucleic acid molecule comprising a nucleotide sequence complementary to a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104.
The nucleic acid molecule of claim 84 wherein said molecule is an antisense oligonucleotide directed to a region of a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104.
An isolated antibody which binds to an epitope on a polypeptide of claim 83.
A method for identifying a compound which binds an ion channel encoded by a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104 comprising the steps of:
- a) contacting said ion channel with a compound;
- and
- c) determining whether said compound binds said ion channel.
A method for identifying a compound which binds a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104 comprising the steps of:
- a) contacting said nucleic acid molecule with a compound;
- and
- b) determining whether said compound binds said nucleic acid molecule.
A method for identifying a compound which modulates the activity of an ion channel encoded by a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104 comprising the steps of:
- a) contacting said ion channel with a compound;
- and
- b) determining whether ion channel activity has been modulated.
A method of screening a human subject to diagnose a disorder affecting the brain or genetic predisposition therefor, comprising the steps of:
- (a) assaying nucleic acid of a human subject to determine a presence or an absence of a mutation altering an amino acid sequence, expression, or biological activity of at least one ion channel that is expressed in the brain, wherein the ion channel comprises an amino acid sequence selected from the group consisting of SEQ ID NO:105 and SEQ ID NO:106, and allelic variants thereof, and wherein the nucleic acid corresponds to a gene encoding the ion channel;
- and
- (b) diagnosing the disorder or predisposition from the presence or absence of said mutation, wherein the presence of a mutation altering the amino acid sequence, expression, or biological activity of the ion channel correlates with an increased risk of developing the disorder.
A kit for screening a human subject to diagnose a mental disorder or a genetic predisposition therefor, comprising, in association:
- (a) an oligonucleotide useful as a probe for identifying polymorphisms in a human ion channel gene, the oligonucleotide comprising 6-50 nucleotides in a sequence that is identical or complementary to a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104, except for one sequence difference selected from the group consisting of a nucleotide addition, a nucleotide deletion, or nucleotide substitution;
- and
- (b) a media packaged with the oligonucleotide, said media containing information for identifying polymorphisms that correlate with a mental disorder or a genetic predisposition therefor, the polymorphisms being identifiable using the oligonucleotide as a probe.
A method of identifying an ion channel allelic variant that correlates with a mental disorder, comprising steps of:
- (a) providing a biological sample comprising nucleic acid from a human patient diagnosed with a mental disorder, or from the patient's genetic progenitors or progeny;
- (b) detecting in the nucleic acid the presence of one or more mutations in an ion channel that is expressed in the brain, wherein the ion channel comprises an amino acid sequence selected from the group consisting of SEQ ID NO:105 and SEQ ID NO:106, and allelic variants thereof, and wherein the nucleic acid includes sequence corresponding to the gene or genes encoding the ion channel;
- wherein the one or more mutations detected indicates an allelic variant that correlates with a mental disorder.
A method to identify compounds useful for the treatment of a disorder, said method comprising the steps of:
- (a) contacting a composition comprising an ion channel encoded by a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104 with a compound suspected of binding said ion channel;
- (b) detecting binding between said ion channel and the compound suspected of binding said ion channel;
- wherein compounds identified as binding said ion channel are candidate compounds useful for the treatment of a disorder.
A method for identifying a compound useful as a modulator of binding between an ion channel encoded by a sequence selected from the group consisting of SEQ ID NO:103 and SEQ ID NO:104 and a binding partner of said ion channel comprising the steps of:
- (a) contacting the binding partner and a composition comprising said ion channel in the presence and in the absence of a putative modulator compound;
- (b) detecting binding between the binding partner and said ion channel;
- wherein decreased or increased binding between the binding partner and said ion channel in the presence of the putative modulator, as compared to binding in the absence of the putative modulator is indicative a modulator compound useful for the treatment of a disorder.
A chimeric receptor comprising at least 5 amino acid residues, said receptor comprising at least a portion of a sequence selected from the group consisting of SEQ ID NO:105 and SEQ ID NOS:106.
Dependencies are inferred from the claim text.