Juvenile hemochromatosis gene (HFE2A), expression products and uses thereof

Abstract

Polynucleotide and polypeptide sequences for HFE2A, as well as mutations associated with juvenile hemochromatosis, and methods of utilizing these for the screening and identification of agents for the treatment of diseases of iron metabolism, including small organic compounds, are disclosed along with methods of treating and/or ameliorating diseases of iron metabolism, especially in human patients are disclosed. Diagnostic compounds, kits and methods using HFE2A are also described.

1 - 117 . (canceled) 
     
     
         118 . An isolated precursor RNAi molecule, comprising first and second oligonucleotide sequences that anneal to each other under biological conditions and wherein at least one of said oligonucleotide sequences is sufficiently complementary to an HFE2A-encoding RNA polynucleotide so as to inhibit expression of an HFE2A polypeptide. 
     
     
         119 . The isolated precursor RNAi molecule of  claim 118 , wherein said at least one of said oligonucleotide sequences is complementary to at least a portion of a sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8 and 9. 
     
     
         120 . The isolated precursor RNAi molecule of  claim 118 , wherein said molecule is capable of inhibiting hemojuvelin activity in a cell. 
     
     
         121 . The isolated precursor RNAi molecule of  claim 118 , wherein said molecule is capable of inhibiting hepcidin production in a cell. 
     
     
         122 . The isolated precursor RNAi molecule of  claim 118 , wherein said molecule is capable of inhibiting iron transport across a membrane in a cell. 
     
     
         123 . A method for inhibiting hemojuvelin activity in a cell, comprising contacting a cell with an isolated precursor RNAi molecule, wherein said molecule comprises first and second oligonucleotide sequences that anneal to each other under biological conditions and wherein at least one of said oligonucleotide sequences is sufficiently complementary to an HFE2A-encoding RNA polynucleotide so as to inhibit expression of an HFE2A polypeptide isolated antisense molecule complementary to an HFE2A polynucleotide, wherein said isolated precursor RNAi molecule inhibits hemojuvelin activity in said cell. 
     
     
         124 . The method of  claim 123 , wherein said at least one of said oligonucleotide sequences is complementary to at least a portion of a sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8 and 9. 
     
     
         125 . The method of  claim 123 , wherein said cell is a member selected from the group consisting of a macrophage, inflammatory cell, Liver cell, intestinal cell, hematopoietic cell, pancreatic cell, skeletal muscle cell, a cell of the nervous system or a Caco2 cell. 
     
     
         126 . The method of  claim 125 , wherein said cell is a Caco2 cell. 
     
     
         127 . The method of  claim 125 , wherein said cell is a macrophage. 
     
     
         128 . The method of  claim 125 , wherein said cell is a hepatocyte. 
     
     
         129 . The method of  claim 125 , wherein said cell is an intestinal cell. 
     
     
         130 . The method of  claim 125 , wherein said cell is a CHO cell. 
     
     
         131 . A method for modulating iron transport across the membrane of a cell, comprising contacting a cell with an isolated precursor RNAi molecule, wherein said molecule comprises first and second oligonucleotide sequences that anneal to each other under biological conditions and wherein at least one of said oligonucleotide sequences is sufficiently complementary to an HFE2A-encoding RNA polynucleotide so as to inhibit expression of an HFE2A polypeptide. isolated antisense molecule complementary to an HFE2A polynucleotide, wherein said isolated precursor RNAi molecule inhibits iron transport across the membrane of said cell. 
     
     
         132 . The method of  claim 131 , wherein said at least one of said oligonucleotide sequences is complementary to at least a portion of a sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8 and 9. 
     
     
         133 . The method of  claim 131 , wherein said modulation of iron transport across a cell membrane results in release of iron by said cell. 
     
     
         134 . The method of  claim 131 , wherein said modulation of iron transport across a cell membrane results in iron uptake by said cell. 
     
     
         135 . The method of  claim 131 , wherein said cell is a member selected from the group consisting of a macrophage, inflammatory cell, Liver cell, intestinal cell, hematopoietic cell, pancreatic cell, skeletal muscle cell, a cell of the nervous system or a Caco2 cell. 
     
     
         136 . The method of  claim 135 , wherein said cell is a Caco2 cell. 
     
     
         137 . The method of  claim 135 , wherein said cell is a macrophage. 
     
     
         138 . The method of  claim 135 , wherein said cell is a hepatocyte. 
     
     
         139 . The method of  claim 135 , wherein said cell is an intestinal cell. 
     
     
         140 . The method of  claim 135 , wherein said cell is a CHO cell. 
     
     
         141 . A method for inhibiting hepcidin production in a cell, comprising contacting a cell with an isolated precursor RNAi molecule, wherein said molecule comprises first and second oligonucleotide sequences that anneal to each other under biological conditions and wherein at least one of said oligonucleotide sequences is sufficiently complementary to an HFE2A-encoding RNA polynucleotide so as to inhibit expression of an HFE2A polypeptide. isolated antisense molecule complementary to an HFE2A polynucleotide, wherein said isolated precursor RNAi molecule inhibits hepcidin production in said cell. 
     
     
         142 . The method of  claim 141 , wherein said at least one of said oligonucleotide sequences is complementary to at least a portion of a sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8 and 9. 
     
     
         143 . The method of  claim 141 , wherein said cell is a member selected from the group consisting of a macrophage, inflammatory cell, Liver cell, intestinal cell, hematopoietic cell, pancreatic cell, skeletal muscle cell, a cell of the nervous system or a Caco2 cell. 
     
     
         144 . The method of  claim 143 , wherein said cell is a Caco2 cell. 
     
     
         145 . The method of  claim 143 , wherein said cell is a macrophage. 
     
     
         146 . The method of  claim 143 , wherein said cell is a hepatocyte. 
     
     
         147 . The method of  claim 143 , wherein said cell is an intestinal cell. 
     
     
         148 . The method of  claim 143 , wherein said cell is a CHO cell.