Viral expression construct comprising a fibroblast growth factor 21 (fgf21) coding sequence

Abstract

The invention relates to a viral expression construct and related viral vector and nucleic acid molecule and composition and to their use wherein said construct and vector are suitable for expression in a mammal and comprise a nucleotide sequence encoding a Fibroblast growth factor 21 (FGF21) to be expressed in liver, adipose tissue and/or skeletal muscle.

1 . A viral expression construct suitable for expression in a mammal and comprising a nucleotide sequence encoding a Fibroblast growth factor 21 (FGF21) to be expressed in liver, adipose tissue and/or skeletal muscle. 
     
     
         2 . The viral expression construct according to  claim 1  comprising a nucleotide sequence encoding a FGF21 suitable for expression in a mammal and at least one of elements a), b), c), d) and e):
 (a) a liver-specific promoter 
 (b) an adipose tissue-specific promoter 
 (c) a combination of an ubiquitous promoter and at least one nucleotide sequence encoding a target sequence of a microRNA expressed in the liver and at least one nucleotide sequence encoding a target sequence of a microRNA expressed in the heart, wherein said combination enables specific expression in adipose tissue 
 (d) a skeletal muscle promoter and 
 (e) a combination of an ubiquitous promoter and an adeno-associated virus (AAV) vector sequence, wherein said combination enables specific expression in skeletal muscle. 
 
     
     
         3 . The viral expression construct according to  claim 1 , wherein the nucleotide sequence encoding a FGF21 suitable for expression in a mammal is selected from the group consisting of:
 (a) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence that has at least 60% sequence identity with the amino acid sequence of SEQ ID NO: 1, 2 or 3   (b) a nucleotide sequence that has at least 60% sequence identity with the nucleotide sequence of SEQ ID NO: 4, 5, 6, 7, 8, 9, 10 or 11   (c) a nucleotide sequence the sequence of which differs from the sequence of a nucleotide sequence of (b) due to the degeneracy of the genetic code.   
     
     
         4 . The viral expression construct according to  claim 2 , wherein the nucleotide sequence encoding a target sequence of a microRNA expressed in the liver and the nucleotide sequence encoding a target sequence of a microRNA expressed in the heart is selected from a group consisting of sequences SEQ ID NO: 12 to 30 and/or combinations thereof. 
     
     
         5 . The viral expression construct according to  claim 1 , wherein the liver-specific promoter is the human α1-antitrypsin (hAAT) promoter and/or the adipose tissue-specific promoter is the mini/aP2 and/or the mini/UCP1 promoter and/or the skeletal muscle promoter is the C5-12 promoter and/or the ubiquitous promoter is the cytomegalovirus (CMV) promoter and/or the CAG promoter. 
     
     
         6 . The viral vector comprising a viral expression construct according to  claim 1 , wherein said viral vector is an adenovirus vector, an adeno-associated virus vector, a retrovirus vector or a lentivirus vector, preferably an adeno-associated virus vector selected from the group consisting of an adeno-associated virus 1 (AAV1) vector, an adeno-associated virus 8 (AAV8) vector, and an adeno-associated virus 9 (AAV9) vector. 
     
     
         7 . A nucleic acid molecule suitable for expression in a mammal and represented by a mammalian codon optimized nucleotide sequence encoding a FGF21 to be expressed in liver, adipose tissue and/or skeletal muscle. 
     
     
         8 . The nucleic acid molecule according to  claim 7 , wherein the nucleotide sequence has at least 70% sequence identity with the nucleotide sequence of SEQ ID NO: 4, 5, 6, 7, 8, 9, 10 or 11. 
     
     
         9 . A composition comprising a viral expression construct as defined in  claim 1 , together with one or more pharmaceutically acceptable excipients or vehicles. 
     
     
         10 .- 16 . (canceled) 
     
     
         17 . A method for preventing, delaying, reverting, curing and/or treating a metabolic disorder comprising the use of a viral expression construct as defined in  claim 1 . 
     
     
         18 . The method according to  claim 17 , wherein the metabolic disorder is diabetes and/or obesity. 
     
     
         19 . The method according to  claim 17 , wherein the metabolic disorder is NASH. 
     
     
         20 . A method for preventing, delaying, reverting, curing and/or treating liver inflammation and/or fibrosis, comprising the use of a viral expression construct as defined in  claim 1 . 
     
     
         21 . A method for extending healthy lifespan, comprising the use of a viral expression construct as defined in  claim 1 . 
     
     
         22 . A method for preventing, delaying, reverting, curing and/or treating cancer, preferably liver cancer, comprising the use of a viral expression construct as defined in  claim 1 . 
     
     
         23 .- 28 . (canceled)