METHODS AND COMPOSITIONS FOR TAU REDUCTION GENE THERAPY
The present disclosure provides methods and compositions for the treatment of tauopathies, such as, for example, Alzheimer's Disease or FTDP-17. The methods and compositions of the present disclosure comprise isolated nucleic acid molecules, rAAV vectors and rAAV viral vectors comprising polynucleotide sequences encoding for artificial micro RNAs (amiRNAs) directed against MAPT.
1 . A recombinant adeno-associated virus (rAAV) vector comprising at least one polynucleotide sequence encoding at least one artificial micro RNA (amiRNA) directed against MAPT, wherein the amiRNA directed against MAPT comprises the nucleic acid sequence set forth in any one of SEQ ID NOs: 139-186.
2 . The rAAV vector of claims 1 , wherein the polynucleotide sequence encoding the at least one amiRNA directed against MAPT comprises the nucleic acid sequence set forth in any one of SEQ ID NOs: 41-88.
3 . An rAAV vector of claim 1 or 2 , wherein the rAAV vector further comprises a first inverted terminal repeat (ITR) sequence, wherein the first ITR sequence is an AAV2 ITR sequence.
4 . The rAAV vector of claim 3 , wherein the first ITR sequence comprises the sequence set forth in SEQ ID NO: 15.
5 . An rAAV vector of any one of claim 1-4 wherein the rAAV vector further comprises a second ITR sequence, wherein the second ITR sequence is an AAV2 ITR sequence.
6 . The rAAV vector of claim 5 , wherein the second ITR sequence comprises the sequence set forth in SEQ ID NO: 16.
7 . The rAAV vector of any one of claims 1-6 , wherein the rAAV vector further comprises a first promoter sequence, wherein the first promoter sequence is a murine U6 promoter sequence.
8 . The rAAV vector of claim 7 , wherein the murine U6 promoter sequence comprises the nucleic acid sequence set forth in SEQ ID NO: 31.
9 . The rAAV vector of any one of claims 1-8 , wherein the rAAV vector further comprises a second promoter sequence, wherein the second promoter sequence is a chicken β-actin hybrid (CBh) promoter sequence.
10 . The rAAV vector of claim 9 , wherein the CBh promoter sequence comprises the nucleic acid sequence set forth in SEQ ID NO: 29.
11 . The rAAV vector of any one of claims 1-10 , wherein the rAAV vector further comprises a termination signal.
12 . The rAAV vector of claim 11 , wherein the termination signal comprises the nucleic acid sequence set forth in SEQ ID NO: 40.
13 . An rAAV vector comprising, in the 5′ to 3′ direction:
a. a first AAV2 ITR sequence;
b. a murine U6 promoter sequence;
c. a polynucleotide sequence encoding for at least one amiRNA directed against MAPT, wherein the amiRNA directed against MAPT comprises the nucleic acid sequence set forth in any one of SEQ ID NOs: 139-186;
d. a termination sequence;
e. a CBh promoter sequence;
f. a synthetic polyA sequence; and
g. a second AAV2 ITR sequence.
14 . The rAAV vector of claim 13 , wherein the rAAV vector comprises the sequence set forth in SEQ ID NO: 14.
15 . An rAAV viral vector comprising:
(a) an AAV capsid protein; and
(b) an rAAV vector of any one of claims 1 - 14 .
16 . The rAAV viral vector of claim 15 , wherein the AAV capsid protein is an AAV9 capsid protein.
17 . A pharmaceutical composition comprising the rAAV vector of any one of claims 1-14 or the rAAV viral vector of claim 15 or 16 ; and at least one pharmaceutically acceptable excipient and/or additive.
18 . A method for treating a tauopathy in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the rAAV vector of any one of claims 1-14 , the rAAV viral vector of claim 15 or 16 , or the pharmaceutical composition of claim 17 .
19 . The method of claim 18 , wherein the tauopathy is Alzheimer's Disease or Frontotemporal Dementia and Parkinsonism Linked To Chromosome 17 (FTDP-17).
20 . The method of claim 18 or 19 , wherein the subject has one or more mutations in the MAPT gene.
21 . The rAAV vector of any one of claims 1-14 , the rAAV viral vector of claim 15 or 16 , or the pharmaceutical composition of claim 17 for use in the treatment of a tauopathy.
22 . The rAAV vector, the rAAV viral vector, or the pharmaceutical compositions for use of claims 21 , wherein the tauopathy is Alzheimer's Disease or FTDP-17.
23 . The rAAV vector, the rAAV viral vector, or the pharmaceutical compositions of claim 21 or 22 , wherein the tauopathy is associated with one or more mutations in the MAPT gene.