Detection of Mutations in a Gene Associated with Resistance to Viral Infection, OAS1

Abstract

A method for detecting a mutation related to the gene encoding OAS1. This and other disclosed mutations correlate with resistance of humans to viral infection including hepatitis C. Also provided is a therapeutic agent consisting of a protein or polypeptide encoded by the mutated gene, or a polynucleotide encoding the protein or polypeptide. Inhibitors of human OAS1, including antisense oligonucleotides, methods, and compositions specific for human OAS1, are also provided.

What is claimed is: 
     
         1 . A human genetic screening method for identifying an oligoadenylate synthetase gene (OAS1) mutation comprising detecting in a nucleic acid sample the presence of an OAS1 point mutation selected from the group consisting of: substitution of a non-reference nucleotide for a reference nucleotide at nucleotide position 2135728, 2135749, 2135978, 2144072, 2144088, 2144116, 2144321, 2131025, 2133961, 2139587, 2144294, 2144985, 2156523, and 2156638 of reference sequence SEQ ID NO:19; and deletion of the reference nucleotide at position 2156595 of reference sequence SEQ ID NO:19; thereby identifying said mutation. 
     
     
         2 . An isolated polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:20-30, SEQ ID NO:32-35, and SEQ ID NO:46-52. 
     
     
         3 . An isolated polypeptide consisting of at least one amino acid sequence selected from the group consisting of SEQ ID NO:75-84 and having at least 80% sequence similarity to a polypeptide selected from the group consisting of SEQ ID NO:20-30, SEQ ID NO:32-35, and SEQ ID NO:46-52. 
     
     
         4 . An isolated polypeptide consisting of at least one amino acid sequence selected from the group consisting of SEQ ID NO:75-84 and having at least 80% sequence similarity to a polypeptide selected from the group consisting of:
 (a) amino acids 219 through 238 of any one of SEQ ID NO:22 and SEQ ID NO:25;   (b) amino acids 231 through 250 of SEQ ID NO:23;   (c) amino acids 347 through 366 of any one of SEQ ID NO: 26-29, SEQ ID NO:33-34, and SEQ ID NO:50;   (d) amino acids 295 through 314 of SEQ ID NO:32;   (e) amino acids 189 through 208 of SEQ ID NO:46;   (f) amino acids 61 through 80 of SEQ ID NO:47.   
     
     
         5 . The polypeptide of any one of  claims 2 - 4  covalently attached to a polypeptide comprising a protein transduction domain. 
     
     
         6 . The polypeptide of  claim 5  wherein the protein transduction domain is comprised of a polypeptide selected from the group consisting of SEQ ID NO:85-94. 
     
     
         7 . The polypeptide of  claim 5  wherein the protein transduction domain is comprised of a polypeptide having at least 80% sequence similarity to a polypeptide selected from the group consisting of SEQ ID NO:85-94. 
     
     
         8 . The polypeptide of  claim 5  wherein the protein transduction domain differs from a polypeptide selected from the group consisting of SEQ ID NO:85-94 by the addition or substitution of an arginine, lysine, or histidine. 
     
     
         9 . The polypeptide of any one of  claims 2 - 8  covalently attached to polyethylene glycol. 
     
     
         10 . The polypeptide of any one of  claims 2 - 8  encapsulated in a liposome. 
     
     
         11 . The polypeptide of any one of  claims 2 - 8  covalently attached to an endosome disrupting agent. 
     
     
         12 . The polypeptide of any one of  claims 2 - 8  noncovalently attached to an endosome disrupting agent. 
     
     
         13 . The polypeptide of  claim 11  or  12  wherein the endosome disrupting agent is pH sensitive. 
     
     
         14 . The polypeptide of any one of  claims 2 - 13  covalently conjugated to a sugar moiety. 
     
     
         15 . The polypeptide of  claim 14  wherein the sugar moiety is comprised of galactose or mannose. 
     
     
         16 . An isolated polypeptide produced by the method comprising:
 (a) expressing the polypeptide of any one of  claims 2 - 8  by a cell; and   (b) recovering said polypeptide.   
     
     
         17 . The polypeptide of any one of  claims 2 - 8  which is produced by a recombinant host cell. 
     
     
         18 . The polypeptide of any one of  claims 2 - 8  comprising a heterologous polypeptide sequence. 
     
     
         19 . A composition comprising the polypeptide of any one of  claims 2 - 18  and a pharmaceutically-acceptable carrier. 
     
     
         20 . An isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide sequence of any one of  claims 2 - 8 . 
     
     
         21 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:31, SEQ ID NO:36-45 and SEQ ID NO:55-56. 
     
     
         22 . A recombinant vector comprising the isolated polynucleotide of any one of  claims 20  and  21 . 
     
     
         23 . An expression vector comprising the isolated polynucleotide of any one of  claims 20  and  21 , operably linked to an expression control sequence. 
     
     
         24 . A host cell transformed or transfected with an expression vector according to  claim 23 . 
     
     
         25 . An expression vector according to  claim 23 , wherein the vector is a viral vector. 
     
     
         26 . An expression vector according to  claim 23 , wherein the vector is a viral vector selected from the group consisting of an adenovirus vector, adeno-associated virus vector, baculovirus, semliki forest virus vector, sindbis virus vector, pox virus vector, vaccinia virus vector, avian poxvirus vector, avipox virus vector, fowlpox virus vector, canarypox virus vector, alphavirus vector, fowlpox virus vector or lentivirus vector. 
     
     
         27 . A composition comprising a first component selected from the group consisting of a pharmaceutically acceptable carrier, and a second component selected from the group consisting of an expression vector according to any one of  claims 23 ,  25  and  26 . 
     
     
         28 . A method of treating viral infection in a mammal comprising administering to a mammal in need of such treatment a composition comprising oligoadenylate synthetase 1. 
     
     
         29 . The method of  claim 28  wherein said oligoadenylate synthetase 1 is the polypeptide of any one of  claims 2 - 8 . 
     
     
         30 . The method of  claim 28  wherein said oligoadenylate synthetase 1 is expressed by the polynucleotide of any one of  claims 20  and  21 . 
     
     
         31 . The method of  claim 28  comprising administering a composition according to any one of  claims 19  and  27 . 
     
     
         32 . The method of  claim 28  wherein said viral infection is an infection with a double-stranded RNA virus. 
     
     
         33 . The method of  claim 28  wherein said viral infection is an infection with a flavivirus. 
     
     
         34 . The method of  claim 33  wherein said flavivirus is the hepatitis C virus. 
     
     
         35 . The method of  claim 28  wherein said viral infection is an infection with a paramyxovirus. 
     
     
         36 . The method of  claim 28  wherein said viral infection is an infection with a virus selected from the group consisting of HIV, respiratory syncytial virus, influenza, coronavirus, parainfluenza, hepatitis A, measles, mumps, West Nile, dengue, yellow fever, polio, herpes, and human papilloma virus. 
     
     
         37 . The method of  claim 28  where said viral infection is severe acute respiratory syndrome. 
     
     
         38 . The method of  claim 28  wherein said mammal does not naturally produce said oligoadenylate synthetase 1. 
     
     
         39 . A method of treating cancer in a mammal comprising administering to a mammal in need of such treatment a composition comprising oligoadenylate synthetase 1. 
     
     
         40 . The method of  claim 39  wherein said oligoadenylate synthetase 1 is the polypeptide of any one of  claims 2 - 8 . 
     
     
         41 . The method of  claim 39  wherein said oligoadenylate synthetase 1 is expressed by one of the polynucleotides of  claims 20  and  21 . 
     
     
         42 . The method of  claim 39  comprising administering a composition from any one of  claims 19  and  27 . 
     
     
         43 . The method of  claim 39  wherein said cancer is prostate cancer. 
     
     
         44 . A monoclonal antibody directed against an epitope on a polypeptide of any one of  claims 2 - 8 . 
     
     
         45 . The antibody of  claim 44 , wherein the antibody is not cross-reactive with another distinct polypeptide of any one of  claims 2 - 8 . 
     
     
         46 . A composition comprising a pharmaceutically acceptable carrier and the antibody selected from any one of  claims 44  and  45 . 
     
     
         47 . An isolated interfering polynucleotide that specifically binds with a target comprising the polynucleotide of any one of  claims 20  and  21 . 
     
     
         48 . The interfering polynucleotide of  claim 47  wherein the target binding site is at least 25 consecutive nucleotides. 
     
     
         49 . A composition comprising the interfering polynucleotide of  claim 47  and a pharmaceutical carrier. 
     
     
         50 . An isolated antisense polynucleotide that specifically binds with SEQ ID NO:19. 
     
     
         51 . A composition comprising the antisense polynucleotide of  claim 50  and a pharmaceutical carrier. 
     
     
         52 . An isolated ribozyme directed at a target comprising the polynucleotide of any one of  claims 20  and  21 . 
     
     
         53 . A composition comprising the ribozyme of  claim 52  and a pharmaceutical carrier. 
     
     
         54 . A method of treating viral infection in a mammal comprising the step of inhibiting oligoadenylate synthetase 1 in a mammal in need of said treatment. 
     
     
         55 . The method of  claim 54  wherein said inhibiting oligoadenylate synthetase 1 comprises administering the composition of any of one of  claims 46 ,  49 ,  51 ,  53 , and  80 . 
     
     
         56 . The method of  claim 54  wherein said oligoadenylate synthetase 1 is the polypeptide of any one of  claims 2 - 8 . 
     
     
         57 . The method of  claim 54  wherein said oligoadenylate synthetase 1 is expressed by the polynucleotide of any one of  claims 20  and  21 . 
     
     
         58 . The method of  claim 54  wherein said viral infection is infection with a virus selected from the group consisting of flavivirus, HIV, respiratory syncytial virus, influenza, coronavirus, parainfluenza, hepatitis A, measles, mumps, West Nile, dengue, yellow fever, polio, herpes, and human papilloma virus. 
     
     
         59 . The method of  claim 54  wherein said viral infection is severe acute respiratory syndrome. 
     
     
         60 . A method of treating insulin dependent diabetes mellitus in a mammal comprising the step of inhibiting oligoadenylate synthetase 1 in a mammal in need of said treatment. 
     
     
         61 . The method of  claim 60  wherein said oligoadenylate synthetase 1 is the polypeptide of any one of  claims 2 - 8 . 
     
     
         62 . The method of  claim 60  wherein said oligoadenylate synthetase 1 is expressed by the polynucleotide of any one of  claims 20  and  21 . 
     
     
         63 . The method of  claim 60  wherein said inhibiting oligoadenylate synthetase 1 comprises administering a composition from any one of  claims 46 ,  49 ,  51 ,  53  and  80 . 
     
     
         64 . The method of  claim 1 , wherein the screening method identifies susceptibility to viral infection in said human. 
     
     
         65 . The method of  claim 1  wherein the screening method identifies predisposition to diabetes mellitus in said human. 
     
     
         66 . The method of  claim 1 , wherein the screening method identifies predisposition to schizophrenia in said human. 
     
     
         67 . The method of  claim 1 , wherein the screening method identifies susceptibility to cancer in said human. 
     
     
         68 . The method of  claim 67 , wherein the cancer is prostate cancer. 
     
     
         69 . The method of  claim 1 , wherein the screening method identifies patient responsiveness to therapeutic treatments for viral infection. 
     
     
         70 . The method of  claim 69  wherein the therapeutic treatment is interferon-based. 
     
     
         71 . The method of  claim 69  wherein patient response is measured by sustained viral clearance. 
     
     
         72 . A method for treating schizophrenia in a human comprising the step of inhibiting oligoadenylate synthetases 1 in a human in need of said treatment. 
     
     
         73 . The method of  claim 72  wherein said oligoadenylate synthetase 1 is the polypeptide of any one of  claims 2 - 8 . 
     
     
         74 . The method of  claim 72  wherein said oligoadenylate synthetase 1 is expressed by the polynucleotide of any one of  claims 20  and  21 . 
     
     
         75 . The method of  claim 72  wherein said inhibiting oligoadenylate synthetase 1 comprises administering a composition from any one of  claims 46 ,  49 ,  51 ,  53  and  80 . 
     
     
         76 . The method of  claim 72  wherein said inhibitor comprises a small molecule drug. 
     
     
         77 . The method of  claim 54  wherein said inhibitor comprises a small molecule drug. 
     
     
         78 . The method of  claim 60  wherein said inhibitor comprises a small molecule drug. 
     
     
         79 . An antibody fragment developed from a monoclonal antibody directed against an epitope on a polypeptide of any one of  claims 2 - 8 . 
     
     
         80 . A composition comprising a pharmaceutically acceptable carrier and the antibody fragment of  claim 79 . 
     
     
         81 . The method of  claim 1 , wherein the screening method identifies susceptibility to hepatitis C infection in said human.