Treatment of hyperbilirubinemia

Abstract

The invention relates to a nucleic acid sequence useful in the treatment of hyperbilirubinemia, in particular in the treatment of Crigler-Najjar syndrome. More particularly, the nucleic acid sequence of the present invention is a codon-optimized UGT1A1 coding sequence.

We claim: 
     
         1 . A nucleic acid sequence which is a codon optimized UGT1A1 coding sequence, wherein the optimized coding sequence has an increased GC content and/or has a decreased number of alternative open reading frames as compared to the wild-type coding sequence. 
     
     
         2 . The nucleic acid sequence according to  claim 1 , comprising a nucleotide sequence at least 80% identical to SEQ ID NO:2 or SEQ ID NO:3. 
     
     
         3 . The nucleic acid sequence according to  claim 2 , comprising the nucleotide sequence shown in SEQ ID NO:2 or SEQ ID NO:3. 
     
     
         4 . A nucleic acid construct comprising the nucleic acid sequence according to  claim 1 . 
     
     
         5 . The nucleic acid construct according to  claim 4 , wherein said nucleic acid construct comprises said nucleic acid sequence operably linked to a promoter is an expression cassette. 
     
     
         6 . The nucleic acid construct according to  claim 5 , wherein the promoter is a liver-specific promoter. 
     
     
         7 . The nucleic acid construct according to  claim 5 , wherein the promoter is selected from the group consisting of the hAAT promoter, alpha-1 antitrypsin promoter (hAAT), the transthyretin promoter, the albumin promoter and the thyroxine-binding globulin (TBG) promoter. 
     
     
         8 . The nucleic acid construct according to  claim 5 , said nucleic acid construct further comprising an intron. 
     
     
         9 . The nucleic acid construct according to  claim 8 , wherein the intron is selected from the group consisting of a human beta globin b2 (HBB2) intron, a FIX intron and a chicken beta-globin intron. 
     
     
         10 . The nucleic acid construct according to  claim 8 , wherein the intron is a modified intron with decreased or no alternative open reading frames (ARFs). 
     
     
         11 . The nucleic acid construct according to  claim 10 , wherein said modified intron is a modified HBB2 intron, a modified FIX intron or a modified chicken beta-globin intron. 
     
     
         12 . The nucleic acid construct according to  claim 11 , wherein said modified intron is the modified HBB2 intron of SEQ ID NO: 6, the modified FIX intron of SEQ ID NO:8, or the modified chicken beta-globin intron of SEQ ID NO:10. 
     
     
         13 . A vector comprising the nucleic acid sequence of  claim 1 . 
     
     
         14 . The vector according to  claim 13 , wherein said vector is a viral vector. 
     
     
         15 . The vector according to  claim 13 , wherein said viral vector is a retroviral vector, a single-stranded or double-stranded self-complementary AAV vector. 
     
     
         16 . The vector according to  claim 14 , wherein the AAV vector has an AAV-derived capsid selected from the group consisting of an AAV-1, -2, -5, -6, -7, -8, -9, -rh10, -rh74 and -dj capsid, or wherein the AAV vector has a chimeric capsid. 
     
     
         17 . The vector according to  claim 15 , wherein the AAV vector has an AAV8 capsid. 
     
     
         18 . The vector according to  claim 15 , wherein the AAV vector is a pseudotyped AAV vector. 
     
     
         19 . A cell transformed with the nucleic acid sequence according to  claim 1 . 
     
     
         20 . The cell according to  claim 19 , wherein said cell is a liver cell or a muscle cell. 
     
     
         21 . A method of treating Crigler-Najjar syndrome type I or II or Gilbert syndrome comprising expression of a nucleic acid sequence according to  claim 1  in a cell or tissue of a subject having Crigler-Najjar syndrome type I or II or Gilbert syndrome. 
     
     
         22 . An intron which is a modified intron with decreased open reading frames. 
     
     
         23 . The intron according to  claim 22 , which is a modified HBB2 intron, a modified FIX intron, or a modified chicken beta-globin intron. 
     
     
         24 . A nucleic acid construct comprising the intron according to  claim 22 . 
     
     
         25 . The nucleic acid construct according to  claim 24 , further comprising a gene of interest and one or more additional expression control sequences. 
     
     
         26 . The nucleic acid construct according to  claim 25 , wherein the said additional expression control sequence is an ubiquitous or tissue-specific promoter. 
     
     
         27 . A vector comprising the intron according to  claim 23 . 
     
     
         28 . The vector according to  claim 27 , which is a viral vector. 
     
     
         29 . The vector according to  claim 28 , wherein said viral vector is a single-stranded or double-stranded self-complementary AAV vector. 
     
     
         30 . The vector according to  claim 29 , wherein the AAV vector has an AAV-derived capsid. 
     
     
         31 . The vector according to  claim 29 , wherein the AAV vector has an AAV8 capsid. 
     
     
         32 . The vector according to  claim 29 , wherein the AAV vector is a pseudotyped AAV vector. 
     
     
         33 . A cell transformed with the nucleic acid construct according to  claim 24 . 
     
     
         34 . A method of gene or cell therapy in a subject comprising expressing a nucleic acid construct according to  claim 25  in a subject in need of gene or cell therapy.