AAV vector for neuronal expression of progranulin
The present invention relates to the field of recombinant viral vectors suitable for the delivery of therapeutic genes in vivo. Described is an adeno-associated virus (AAV) vector comprising (i) a human growth hormone intron 3 (hGHi3) sequence (ii) a synapsin promoter sequence and/or (iii) a progranulin 3′ untranslated region (UTR) sequence, operably coupled to a polynucleotide sequence encoding a polypeptide of interest. Specific use of such a vector lies in the enhanced expression of a polypeptide of interest, such as progranulin (PGRN), to treat subjects who have a genetic mutation or intrinsic polypeptide level that is below a physiologically normal level.
1 . An adeno-associated virus (AAV) vector comprising a nucleic acid comprising:
(i) a human growth hormone intron 3 (hGHi3) sequence, wherein the hGHi3 sequence comprises the nucleic acid sequence of SEQ ID NO: 7;
(ii) a synapsin promoter sequence;
(iii) a polynucleotide coding sequence encoding progranulin (PGRN) and comprising a 5′ exonic splicing element (ESE); and
(iv) a PGRN 3′ untranslated region (UTR) sequence.
2 . The AAV vector according to claim 1 , wherein
the AAV vector is of serotype AAV9.
3 . The AAV vector according to claim 1 , wherein the ESE is part of, or inserted into, a 5′ flanking sequence.
4 . The AAV vector according to claim 1 , wherein the ESE is part of, or inserted into, a guide sequence.
5 . The AAV vector according to claim 1 , wherein the ESE is part of a 5′ flanking sequence derived from a wild-type polynucleotide sequence.
6 . The AAV vector according to claim 3 , wherein the 5′ flanking sequence is a 5′ guide sequence derived from wild-type granulin (GRN).
7 . The AAV vector according to claim 6 , wherein the wild-type GRN 5′ guide sequence comprises 350 to 450 base pairs.
8 . The AAV vector according to claim 1 , wherein the polynucleotide sequence comprises the sequence of SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.
9 . A pharmaceutical composition or medicament comprising an AAV vector as claimed in claim 1 and one or more pharmaceutically or physiologically acceptable carriers, excipients, and/or diluents.
10 . A method of treating a neurological disorder in a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition or medicament of claim 9 .
11 . The method of claim 10 , wherein the neurological disorder is frontotemporal dementia (FTD), neuronal ceroid lipofuscinosis (NCL11), amyotrophic lateral sclerosis (ALS), Huntington's disease, Parkinson's disease, or Alzheimer's disease.
12 . The method of claim 10 , wherein the subject: (i) is heterozygous, homozygous or compound heterozygous for GRN mutations, (ii) is suffering from sporadic neurological disease, and/or (iii) has PGRN levels below a physiologically normal level.
13 . The AAV vector according to claim 1 , wherein:
a) the polynucleotide sequence is codon-optimized for expression in humans; or
b) the ESE is upstream of the polynucleotide coding sequence.