TDP-43-binding polypeptides useful for the treatment of neurodegenerative diseases
Provided herein are antigen-binding constructs such as antibodies that bind to the RRM-1 domain of TDP-43. The antigen-binding constructs are capable of blocking the interaction of TDP-43 with NF-κB in cells. Also provided herein are method of using the antigen-binding constructs in the treatment of diseases associated with TPD-43 proteinopathy, such as amyotrophic lateral sclerosis (ALS), frontotemperal lobar degeneration (FTLD), Lewy body disease and motor neuron disease.
1. An antigen-binding construct or a pharmaceutical composition comprising said antigen-binding construct and a pharmaceutically acceptable excipient wherein said antigen-binding construct specifically binds a TAR-DNA binding protein 43 kDa (TDP-43) and comprises at least one heavy chain variable region VH and at least one variable light chain region (VL) and wherein said antigen-binding construct comprises three VH complementarity determining regions (CDRs) and three VL CDRs selected from the group consisting of:
(a) the CDR1 (SEQ ID NO. 7), CDR2 (SEQ ID NO. 8) and CDR3 (SEQ ID NO. 9), of E6_VH1 and the CDR1 (SEQ ID NO. 13), CDR2 (SEQ ID NO. 14) and CDR3 (SEQ ID NO. 15) of E_6Vκ9;
(b) the CDR1 (SEQ ID NO. 10), CDR2 (SEQ ID NO. 11) and CDR3 (SEQ ID NO12) of E6_VH7 and the CDR1 (SEQ ID NO. 13), CDR2 (SEQ ID NO. 14) and CDR3 (SEQ ID NO. 15) of E_6Vκ9;
(c) the CDR1 (SEQ ID NO16), CDR2 (SEQ ID NO. 17) and CDR3 (SEQ ID NO. 18) of C10_VH3, and the CDR1 (SEQ ID NO. 22), CDR2 (SEQ ID NO. 23 and CDR3 (SEQ ID NO. 24) of C10_Vκ3; and
(d) the CDR1 (SEQ ID NO19), CDR2 (SEQ ID NO. 20) and CDR3 (SEQ ID NO. 21) of C10_VH4 and the CDR1 (SEQ ID NO. 22), CDR2 (SEQ ID NO. 23 and CDR3 (SEQ ID NO. 24) of C10_Vκ3.
2. The antigen-binding construct of claim 1 , comprising:
i) the VH of E6_VH7 (SEQ ID NO. 2) and the VL of E6_Vκ9 (SEQ ID NO. 3);
ii) the VH of E6_VH1 (SEQ ID NO. 1) and the VL of E6_Vκ9 (SEQ ID NO. 3);
iii) the VH of C10_VH3 (SEQ ID NO. 4) and the VL of C10_Vκ3 (SEQ ID NO. 6); or
iv) the VH of C10_VH4 (SEQ ID NO. 5) and the VL of C10_Vκ3 (SEQ ID NO. 6).
3. The antigen-binding construct of claim 1 , wherein the construct comprises a peptide linker between the VH and VL, optionally wherein the linker comprises the amino acid sequence SSGGGGSGGGGSGGGGS (SEQ ID NO:47).
4. The antigen-binding construct of claim 1 , wherein the construct comprises E6_Vh7Vκ9 (SEQ ID NO. 28), E6_Vh1Vκ9 (SEQ ID NO. 26), C10_VH3Vκ3 (SEQ ID NO. 30) or C10_VH4Vκ3 (SEQ ID NO. 32).
5. The antigen-binding construct of claim 1 , wherein the construct comprises a secretory signal peptide.
6. The antigen-binding construct of claim 1 , wherein the construct:
a) has an scFv format, an Fab format, a single domain antibody format or an (Fab′) 2 format;
b) comprises an Fc domain; or
c) is humanized.
7. A method of treating a disease characterized by TDP-43 proteinopathy selected from the group consisting of amyotrophic lateral sclerosis (ALS), Alzheimer's disease, motor neuron disease, Parkinson's disease, frontotemperal lobar degeneration (FTLD), mild cognitive impairment (MCI), Lewy body disease, brain trauma and cerebral ischemia in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of:
a) the antigen-binding construct of claim 1 ; or
b) an adeno-associated viral (AAV) vector comprising at least one nucleic acid sequence that encodes at least one antigen-binding construct of claim 1 .
8. A method of producing the antigen-binding construct of claim 1 comprising culturing a host cell under conditions suitable for expressing the antigen-binding construct, wherein the host cell comprises a polynucleotide encoding the antigen binding construct of claim 1 , and purifying the construct.
9. An polynucleotide or set of isolated polynucleotides comprising at least one nucleic acid sequence that encodes at least one of the antigen-binding construct of claim 1 ; a vector or set of vectors comprising one or more of said polynucleotides or sets of polynucleotides; an isolated cell comprising said polynucleotide or set of polynucleotides; or a kit comprising the antigen-binding construct of claim 1 and instructions for use.
10. The antigen-binding construct of claim 1 , wherein the construct comprises a peptide linker between the VH and VL, wherein the linker comprises the amino acid sequence SSGGGGSGGGGSGGGGS (SEQ ID NO:47).
11. The antigen-binding construct of claim 1 , wherein the construct comprises a secretory signal peptide which is MGDNDIHFAFLSTGVHSQVQ (SEQ ID NO:48).