Zinc finger protein compositions for modulation of huntingtin (Htt)
The present disclosure is in the field of modulation of Htt gene expression and HTT protein levels.
1. A zinc finger protein transcription factor (ZFP-TF) repressor comprising a repression domain and a zinc finger protein (ZFP) as follows:
(a) a ZFP designated ZFP-A comprising the amino
acid sequence
(SEQ ID NO: 64)
MAPKKKRKVGIHGVPAAMAERPFQCRICMRNFSQSSDLSRHIRTHTGEKP
FACDICGRKFAQWSTRKRHTKIHTGSQKPFQCRICMRNFSQSSDLSRHIR
THTGEKPFACDICGRKFAQWSTRKRHTKIHTGEKPFQCRICMRKFAQSGD
LTRHTKIHLRQKDAARGSGMDAKSLTAWSRTLVTFKDVFVDFTREEWKLL
DTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEPWLVEREIHQ
ETHPDSETAFEIKSSV;
(b) a ZFP designated ZFP-B comprising the amino
acid sequence
(SEQ ID NO: 65)
MAPKKKRKVGIHGVPAAMAERPFQCRICMRNFSRSDNLSEHIRTHTGEKP
FACDICGRKFAKRCNLRCHTKIHTHPRAPIPKPFQCRICMRNFSQSGDLT
RHIRTHTGEKPFACDICGRKFAQSGDLTRHTKIHTPNPHRRTDPSHKPFQ
CRICMRNFSRSDNLSEHIRTHTGEKPFACDICGRKFAKRCNLRCHTKIHL
RQKDAARGSGMDAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYR
NVMLENYKNLVSLGYQLTKPDVILRLEKGEEPWLVEREIHQETHPDSETA
FEIKSSV;
(c) a ZFP designated ZFP-C comprising the amino
acid sequence
(SEQ ID NO: 66)
MAPKKKRKVGIHGVPAAMAERPFQCRICMRNFSRSDNLSEHIRTHTGEKP
FACDICGRKFAKRCNLRCHTKIHTGSQKPFQCRICMRNFSRSDNLSEHIR
THTGEKPFACDICGRKFAKRCNLRCHTKIHTGSQKPFQCRICMRNFSRSD
NLSEHIRTHTGEKPFACDICGRKFAKRCNLRCHTKIHLRQKDAARGSGMD
AKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVS
LGYQLTKPDVILRLEKGEEPWLVEREIHQETHPDSETAFEIKSSV;
and
(d) a ZFP designated ZFP-D comprising the amino
acid sequence
(SEQ ID NO: 67)
MAPKKKRKVGIHGVPAAMAERPFQCRICMRNFSRSDNLSEHIRTHTGEKP
FACDICGRKFAKRCNLRCHTKIHTHPRAPIPKPFQCRICMRNFSQSSDLS
RHIRTHTGEKPFACDICGRKFAQWSTRKRHTKIHTGEKPFQCRICMRKFA
QSGDLTRHTKIHLRQKDAARGSGMDAKSLTAWSRTLVTFKDVFVDFTREE
WKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEPWLVER
EIHQETHPDSETAFEIKSSV.
2. A polynucleotide encoding the ZFP-TF repressor of claim 1 .
3. The polynucleotide of claim 2 comprising an mRNA encoding the ZFP-TF.
4. An AAV vector comprising the polynucleotide of claim 2 .
5. A pharmaceutical composition comprising the polynucleotide of claim 3 .
6. A pharmaceutical composition comprising the AAV vector of claim 4 .
7. A cell comprising the ZFP-TF repressor of claim 1 .
8. The cell of claim 7 , wherein the cell is a neuron or a fibroblast.
9. A method of reducing apoptosis and increasing ATP levels in a neuron from a subject with Huntington's Disease (HD), the method comprising administering the AAV vector of claim 4 the neuron.
10. A method of reducing motor deficits in a subject with Huntington's Disease (HD), the method comprising administering the AAV vector of claim 4 the striatum of the subject.
11. A method of reducing HTT inclusions in a subject with Huntington's Disease (HD), the method comprising administering the AAV vector of claim 4 the striatum of the subject.
12. The method of claim 11 , wherein the HTT inclusions are perinuclear or EM48+ inclusions.
13. A method of restoring translational biomarkers in a subject with Huntington's Disease (HD), the method comprising administering the AAV vector of claim 4 the striatum of the subject.
14. The method of claim 13 , wherein the biomarkers are PDE10A and/or D1 or D2 receptors.
15. A method of treating and/or preventing Huntington's Disease in a subject in need thereof, the method comprising administering the pharmaceutical composition of claim 5 to the subject in need thereof.
16. A method of treating and/or preventing Huntington's Disease in a subject in need thereof, the method comprising administering the pharmaceutical composition of claim 6 the subject in need thereof.