Mutations in ion channels

Abstract

A method of identifying a subject predisposed to a disorder associated with ion channel dysfunction, comprising ascertaining whether at least one of the genes encoding ion channel subunits in said subject has undergone a mutation event such that a cDNA derived from said subject has the sequence set forth in one of SEQ ID NOS: 1-134.

1 . A method of identifying a subject predisposed to a disorder associated with ion channel dysfunction, comprising ascertaining whether at least one of the genes encoding ion channel subunits in said subject has undergone a mutation event such that a cDNA derived from said subject has the sequence set forth in one of SEQ ID NOS: 1-134.  
     
     
         2 . A method as claimed in  claim 1 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce an epilepsy phenotype in said subject.  
     
     
         3 . A method as claimed in  claim 1 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce one or more disorders associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness and total color-blindness in said subject.  
     
     
         4 . A method as claimed in  claim 1 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce an epilepsy phenotype when expressed in combination with one or more additional mutations or variations in said ion channel subunit genes.  
     
     
         5 . A method as claimed in  claim 1 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce one or more disorders associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness and total color-blindness, when expressed in combination with one or more additional mutations or variations in said ion channel subunit genes.  
     
     
         6 . An isolated nucleic acid molecule encoding a mutant or variant ion channel subunit wherein a mutation event has occurred such that a cDNA derived therefrom has the sequence set forth in one of SEQ ID NOS: 1-134.  
     
     
         7 . An isolated nucleic acid molecule encoding a mutant or variant ion channel subunit as claimed in  claim 6 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce an epilepsy phenotype.  
     
     
         8 . An isolated nucleic acid molecule encoding a mutant or variant ion channel subunit as claimed in  claim 6 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce one or more disorders associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness and total color-blindness.  
     
     
         9 . An isolated nucleic acid molecule encoding a mutant or variant ion channel subunit as claimed in  claim 6 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce an epilepsy phenotype when expressed in combination with one or more additional mutations or variations in said ion channel subunit genes.  
     
     
         10 . An isolated nucleic acid molecule encoding a mutant or variant ion channel subunit as claimed in  claim 6 , wherein said mutation event disrupts the functioning of an assembled ion channel so as to produce one or more disorders associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness and total color-blindness, when expressed in combination with one or more additional mutations or variations in said ion channel subunit genes.  
     
     
         11 . An isolated nucleic acid molecule comprising any one of the nucleotide sequences set forth in SEQ ID NOS: 1-134.  
     
     
         12 . An isolated nucleic acid molecule consisting of any one of the nucleotide sequences set forth in SEQ ID NOS: 1-134.  
     
     
         13 . Canceled.  
     
     
         14 . Canceled.  
     
     
         15 . Canceled.  
     
     
         16 . Canceled.  
     
     
         17 . Canceled.  
     
     
         18 . Canceled.  
     
     
         19 . Canceled.  
     
     
         20 . Canceled.  
     
     
         21 . An expression vector comprising a nucleic acid molecule as claimed in  claim 6  or  claim 77 .  
     
     
         22 . A cell comprising one or more nucleic acid molecules claimed in  claim 6  or  claim 77 .  
     
     
         23 . Canceled.  
     
     
         24 . A cell comprising at least one ion channel type, wherein the or each ion channel type incorporates at least one mutant polypeptide encoded by a nucleic acid molecule as claimed in  claim 6  or  claim 77 .  
     
     
         25 . Canceled.  
     
     
         26 . Canceled.  
     
     
         27 . A method of preparing a polypeptide, comprising the steps of: 
 (1) culturing cells as claimed in either one of claims  22  or  claim 24  under conditions effective for polypeptide production; and    (2) harvesting the polypeptide.    
     
     
         28 . A polypeptide prepared by the method of  claim 27 .  
     
     
         29 . Canceled.  
     
     
         30 . Canceled.  
     
     
         31 . Canceled.  
     
     
         32 . Canceled.  
     
     
         33 . Canceled.  
     
     
         34 . Canceled.  
     
     
         35 . Canceled.  
     
     
         36 . Canceled.  
     
     
         37 . Canceled.  
     
     
         38 . Canceled.  
     
     
         39 . Canceled.  
     
     
         40 . Canceled.  
     
     
         41 . Canceled.  
     
     
         42 . Canceled.  
     
     
         43 . Canceled.  
     
     
         44 . Canceled.  
     
     
         45 . Canceled.  
     
     
         46 . Canceled.  
     
     
         47 . Use of a nucleic acid molecule as claimed in  claim 6  or  claim 77  for the screening of candidate pharmaceutical agents.  
     
     
         48 . Use as claimed in  claim 47  for the screening of candidate pharmaceutical agents useful for the treatment of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness.  
     
     
         49 . Canceled.  
     
     
         50 . Use of a polypeptide as claimed in either  claim 28  or  claim 96  for the screening of candidate pharmaceutical agents.  
     
     
         51 . Use as claimed in  claim 50  for the screening of candidate pharmaceutical agents useful for the treatment of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness.  
     
     
         52 . Canceled.  
     
     
         53 . Use of a cell as claimed in claims  22  or  24  for the screening of candidate pharmaceutical agents.  
     
     
         54 . Use as claimed in  claim 53  for the screening of candidate pharmaceutical agents useful for the treatment of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness.  
     
     
         55 . Canceled.  
     
     
         56 . Canceled.  
     
     
         57 . Canceled.  
     
     
         58 . A genetically modified non-human animal comprising one or more nucleic acid molecules as claimed in  claim 6  or  claim 77 .  
     
     
         59 . Canceled.  
     
     
         60 . Canceled.  
     
     
         61 . Canceled.  
     
     
         62 . Canceled.  
     
     
         63 . Use of a genetically modified non-human animal as claimed in  claim 58  in the screening of candidate pharmaceutical compounds.  
     
     
         64 . Use as claimed in  claim 63  in the screening of candidate pharmaceutical compounds useful in the treatment of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness.  
     
     
         65 . Canceled.  
     
     
         66 . The use of a nucleic acid molecule as claimed in  claim 6  or  claim 77  for the diagnosis of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total colour-blindness.  
     
     
         67 . Canceled.  
     
     
         68 . The use of a polypeptide as claimed in  claim 28  or  claim 96  in the diagnosis of epilepsy or a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness.  
     
     
         69 . Canceled.  
     
     
         70 . Canceled.  
     
     
         71 . Canceled.  
     
     
         72 . A method for the diagnosis of epilepsy comprising the steps of: 
 (1) obtaining DNA from a subject; and    (2) comparing the DNA of one or more subunits of ion channels from said subject to the DNA of the corresponding native subunits;    wherein identification of one or more DNA molecules as claimed in  claim 6  or  claim 77  is an indication of epilepsy, or a predisposition thereto.    
     
     
         73 . A method for the diagnosis of a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total color-blindness, comprising the steps of: 
 (1) obtaining DNA from a subject; and    (2) comparing the DNA of one or more subunits of ion channels from said subject to the DNA of the corresponding native subunits;    wherein identification of one or more DNA molecules as claimed in  claim 6  or  claim 77  is an indication of the disorder, or a predisposition thereto.    
     
     
         74 . Canceled.  
     
     
         75 . Canceled.  
     
     
         76 . Canceled.  
     
     
         77 . An isolated nucleic acid molecule encoding a mutant subunit of a mammalian nicotinic acetylcholine receptor (nAChR), wherein a mutation event selected from the group consisting of point mutations, deletions, insertions and rearrangements has occurred in the nucleotides outside of the M2 domain of the subunit of said mammalian nicotinic acetylcholine receptor, so as to produce an epilepsy phenotype or so as to produce a disorder associated with ion channel dysfunction, including but not restricted to, hyper- or hypo-kalemic Periodic paralysis, myotonias, malignant hyperthermia, myasthenia, cardiac arrhythmias, episodic ataxia, migraine, Alzheimer's disease, Parkinson's disease, schizophrenia, hyperekplexia, anxiety, depression, phobic obsessive symptoms, neuropathic pain, inflammatory pain, chronic/acute pain, Bartter's syndrome, polycystic kidney disease, Dent's disease, hyperinsulinemic hypoglycemia of infancy, cystic fibrosis, congenital stationary night blindness or total colour-blindness.  
     
     
         78 . Canceled.  
     
     
         79 . An isolated nucleic acid molecule as claimed in  claim 77  wherein said mutant subunit is the CHRNA4 subunit.  
     
     
         80 . An isolated nucleic acid molecule as claimed in  claim 78  wherein said mutation event takes place in exon 5.  
     
     
         81 . An isolated nucleic acid molecule as claimed in  claim 79  wherein said mutation event is selected from the group consisting of a C to T nucleotide substitution at position 1006, a G to A nucleotide substitution at position 1106 and a C to G nucleotide substitution at position 1421.  
     
     
         82 . An isolated nucleic acid molecule as claimed in  claim 77  wherein said mutant subunit is the CHRNB2 subunit.  
     
     
         83 . An isolated nucleic acid molecule as claimed in  claim 81  wherein said mutation event takes place in exon 2 or exon 5.  
     
     
         84 . An isolated nucleic acid molecule as claimed in  claim 82  wherein said mutation event is selected from the group consisting of a C to T nucleotide substitution at position 77, a C to G nucleotide substitution at position 901, a T to C nucleotide substitution at position 923 and a G to A nucleotide substitution at position 1235.  
     
     
         85 . Canceled.  
     
     
         86 . Canceled.  
     
     
         87 . Canceled.  
     
     
         88 . Canceled.  
     
     
         89 . Canceled.  
     
     
         90 . Canceled.  
     
     
         91 . Canceled.  
     
     
         92 . Canceled.  
     
     
         93 . Canceled.  
     
     
         94 . Canceled.  
     
     
         95 . Canceled.  
     
     
         96 . Canceled.  
     
     
         97 . A polypeptide encoded by a nucleic acid molecule as claimed in  claim 6  or  77 .