Insulin-like growth factor binding protein-like 1 (igfbpl1) compositions and methods of use thereof

Abstract

The present disclosure relates to insulin-like growth factor binding protein-like 1 (IGFBPL1) compositions and methods of use thereof, including inhibiting SORT1 and DCC activity expressed on neurons and microglia.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 510,577, filed June 27, 2023, the entire contents of which are incorporated herein by reference. Background Technology

[0003] Insulin-like growth factor binding protein-like 1 (IGFBPL1) is a multidomain protein that has been reported to have beneficial activities in neurodegenerative and neuroinflammatory conditions, such as glaucoma, by promoting neurite growth, for example. However, the underlying mechanisms and activities leading to IGFBPL1 activity under such conditions are unknown. Summary of the Invention

[0004] In some aspects, this disclosure provides a fusion protein comprising a half-life extension domain and a human IGFBPL1 (hIGFBPL1) polypeptide or a fragment thereof, wherein the half-life extension domain is operatively linked to the hIGFBPL1 polypeptide or a fragment thereof with or without a linker.

[0005] In some aspects, this disclosure provides a fusion protein comprising a half-life extension domain and a DCC-binding component, wherein the half-life extension domain is operatively linked to the component with or without a linker.

[0006] In some aspects, this disclosure provides a fusion protein comprising a half-life extension domain and a SORT1 binding component, wherein the half-life extension domain is operatively linked to the component with or without a linker.

[0007] In some aspects, this disclosure provides a fusion protein comprising a half-life extension domain and a component for binding SORT1 and DCC, wherein the half-life extension domain is operatively linked to the component with or without a linker.

[0008] In some embodiments, the hIGFBPL1 polypeptide or a fragment thereof comprises a thumb domain, an N-terminal domain, a Kazal domain and an Ig-like domain, a C-terminal domain, or any combination thereof. In some embodiments, the thumb domain comprises the amino acid sequence of SEQ ID NO:25, the N-terminal domain comprises the amino acid sequence of SEQ ID NO:2, the Kazal domain comprises the amino acid sequence of SEQ ID NO:27, the Ig-like domain comprises the amino acid sequence of SEQ ID NO:26, and / or the C-terminal domain comprises the amino acid sequence of SEQ ID NO:3.

[0009] In some embodiments, the hIGFBPL1 fragment does not contain the hIGFBPL1 thumb domain. In some embodiments, the hIGFBPL1 fragment does not contain the hIGFBPL1 N-terminal domain. In some embodiments, the hIGFBPL1 fragment does not contain the hIGFBPL1 Kazal domain. In some embodiments, the hIGFBPL1 fragment does not contain the hIGFBPL1 Ig-like domain. In some embodiments, the hIGFBPL1 fragment does not contain the hIGFBPL1 C-terminal domain.

[0010] In some embodiments, the hIGFBPL1 fragment includes the N-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID NO:2, 158, and 160-166.

[0011] In some embodiments, the hIGFBPL1 fragment includes the C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:179.

[0012] In some embodiments, the hIGFBPL1 fragment includes the thumb domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO: 25.

[0013] In some embodiments, the hIGFBPL1 fragment includes the Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes an amino acid sequence selected from SEQ ID No: 26, 167, and 169.

[0014] In some embodiments, the hIGFBPL1 fragment contains the Kazal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment contains the amino acid sequence of SEQ ID NO:27 or SEQ ID NO:178.

[0015] In some embodiments, the hIGFBPL1 fragment includes the Ig-like domain and C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO:168 or SEQ ID NO:170.

[0016] In some embodiments, the hIGFBPL1 fragment includes the Kazal domain and Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO: 171 or SEQ ID NO: 173.

[0017] In some embodiments, the hIGFBPL1 fragment includes the Kazal domain, Ig-like domain, and C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO: 172 or SEQ ID NO: 174.

[0018] In some embodiments, the hIGFBPL1 fragment includes the N-terminal domain and Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO: 175.

[0019] In some embodiments, the hIGFBPL1 fragment includes the N-terminal domain, Ig-like domain, and C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment. In some embodiments, the hIGFBPL1 fragment includes the amino acid sequence of SEQ ID NO: 176 or SEQ ID NO: 177.

[0020] In some embodiments, the hIGFBPL1 peptide or fragment of hIGFBPL1 comprises one or more domains of mouse IGFBPL1 (mIGFBPL1). In some embodiments, the hIGFBPL1 peptide or fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID No: 151-153 and 155-157.

[0021] In some embodiments, the hIGFBPL1 polypeptide or a fragment thereof comprises the natural signal peptide of hIGFBPL1. In some embodiments, the natural signal peptide comprises the amino acid sequence of SEQ ID NO: 28. In some embodiments, the hIGFBPL1 polypeptide or a fragment thereof comprises a heterologous signal peptide. In some embodiments, the heterologous signal peptide comprises the amino acid sequence of SEQ ID NO: 36.

[0022] In some embodiments, the hIGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID NO:1, 10 and 11, or a fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID NO:37-50.

[0023] In some embodiments, the hIGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID NO:1, 10, and 11, or a fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID NO:37-50, 158, and 160-180.

[0024] In some embodiments, the half-life extension domain is located at the N-terminus of the hIGFBPL1 peptide or a fragment thereof. In some embodiments, the half-life extension domain is located at the C-terminus of the hIGFBPL1 peptide or a fragment thereof.

[0025] In some implementations, the half-life extension domain is selected from Fc domains, albumin domains, transferrin domains, C-terminal peptides, elastin-like peptides, inaccurate repeating peptide sequences, proline-alanine-serine polymers, repeating sequences of glycine-rich peptides, and gelatin-like proteins.

[0026] In some embodiments, the half-life extension domain is an Fc domain. In some embodiments, the Fc domain comprises an amino acid sequence selected from SEQ ID NO: 4-9. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 7. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 9.

[0027] In some embodiments, the half-life extension domain is an albumin polypeptide. In some embodiments, the albumin polypeptide comprises the amino acid sequence of SEQ ID NO: 181.

[0028] In some embodiments, the half-life extension domain and the hIGFBPL1 peptide or a fragment thereof are operably linked without a linker. In some embodiments, the half-life extension domain and the hIGFBPL1 peptide or a fragment thereof are operably linked with a linker. In some embodiments, the linker is a peptide linker. In some embodiments, the linker is a TEV linker or a GS linker. In some embodiments, the linker comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NO: 12-14. In some embodiments, the linker comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NO: 12-14, 23-24, 159, and 182.

[0029] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO: 15-22, 109-110, 134, 138-140, 142, and 148-150. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 15-22, 109-110, 134, 138-140, 142, and 148-150.

[0030] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO:51-72, 111-133, and 183. In some embodiments, the fusion protein comprises the amino acid sequence selected from SEQ ID NO:51-72, 111-133, and 183.

[0031] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO: 94-100 and 102-108.

[0032] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID No: 144-147. In some embodiments, the fusion protein comprises the amino acid sequence selected from SEQ ID NO: 144-147.

[0033] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 15. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 15.

[0034] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 16. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 16.

[0035] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 17. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 17.

[0036] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 18. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 18.

[0037] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 19. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 19.

[0038] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 20. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 20.

[0039] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 21. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 21.

[0040] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence of SEQ ID NO: 22. In some embodiments, the fusion protein comprises the amino acid sequence of SEQ ID NO: 22.

[0041] In some embodiments, the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO: 51-72. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 51-72.

[0042] In some embodiments, hIGFBPL1 or a fragment thereof contains at least one glycosylation mutation. In some embodiments, the glycosylation mutation is located at amino acid residues N166 and / or S268 as identified in SEQ ID NO: 1. In some embodiments, the glycosylation mutation is N166Q, N166D, S268A, or any combination thereof. In some embodiments, the hIGFBPL1 polypeptide containing the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NO: 87-91.

[0043] In some embodiments, the Fc domain contains a glycosylation mutation. In some embodiments, the glycosylation mutation in the Fc domain is at amino acid residue N297, as identified in any of SEQ ID NO: 4-9. In some embodiments, the glycosylation mutation in the Fc domain is N297D or N297Q. In some embodiments, the Fc domain containing the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NO: 92 and 93.

[0044] In some aspects, this disclosure provides a fusion protein dimer comprising two of the fusion proteins disclosed herein. In some embodiments, the two fusion proteins are identical. In some embodiments, the two fusion proteins are different.

[0045] In some aspects, this disclosure provides a fusion protein dimer comprising (i) a first fusion protein comprising a first half-life extension domain operatively linked to a first hIGFBPL1 polypeptide or a fragment thereof with or without a linker, and (ii) a second fusion protein comprising a second half-life extension domain operatively linked to a second hIGFBPL1 polypeptide or a fragment thereof with or without a linker.

[0046] In some embodiments, the first and second IGBBPL1 peptides or fragments thereof are identical. In other embodiments, the first and second IGBBPL1 peptides or fragments thereof are different.

[0047] In some embodiments, the first and second half-life extension domains are identical. In some embodiments, the first and second half-life extension domains are different. In some embodiments, the first half-life extension domain is operatively linked to the N-terminus of a first hIGFBPL1 peptide or a fragment thereof. In some embodiments, the first half-life extension domain is operatively linked to the C-terminus of a first hIGFBPL1 peptide or a fragment thereof. In some embodiments, the second half-life extension domain is operatively linked to the N-terminus of a second hIGFBPL1 peptide or a fragment thereof. In some embodiments, the second half-life extension domain is operatively linked to the C-terminus of a second hIGFBPL1 peptide or a fragment thereof.

[0048] In some embodiments, the first fusion protein includes a third hIGFBPL1 polypeptide or a fragment thereof operatively linked to the half-life extension domain, with or without a linker. In some embodiments, the first and third hIGFBPL1 polypeptides or fragments thereof are identical. In some embodiments, the first and third hIGFBPL1 polypeptides or fragments thereof are different. In some embodiments, the third hIGFBPL1 polypeptide or fragment thereof is operatively linked to the opposite end of the half-life extension domain, compared to the first hIGFBPL1 polypeptide or fragment thereof.

[0049] In some embodiments, the second fusion protein includes a fourth hIGFBPL1 polypeptide or a fragment thereof operatively linked to the half-life extension domain, with or without a linker. In some embodiments, the second and fourth hIGFBPL1 polypeptides or fragments thereof are identical. In some embodiments, the second and fourth hIGFBPL1 polypeptides or fragments thereof are different. In some embodiments, the fourth hIGFBPL1 polypeptide or fragment thereof is operatively linked to the opposite end of the half-life extension domain compared to the second hIGFBPL1 polypeptide or fragment thereof.

[0050] In some embodiments, the first, second, third, and / or fourth hIGFBPL1 polypeptide or fragments thereof comprise amino acid sequences selected from SEQ ID NO:1, 10-11, and 37-50. In some embodiments, the first, second, third, and / or fourth hIGFBPL1 polypeptide or fragments thereof comprise amino acid sequences selected from SEQ ID NO:1, 10-11, 37-50, 158, and 160-180.

[0051] In some embodiments, the first half-life extension domain is a first Fc domain, and the second half-life extension domain is a second Fc domain. In some embodiments, the first and second Fc domains contain mutations for dimerization. In some embodiments, the first Fc domain contains a club-shaped mutation and the second Fc domain contains a mortar-shaped mutation to dimerize the first and second Fc domains. In some embodiments, the first Fc domain contains a mortar-shaped mutation and the second Fc domain contains a club-shaped mutation to dimerize the first and second Fc chains. In some embodiments, the first and second Fc domains each contain an amino acid sequence selected from SEQ ID NO:4-9, 92, and 93.

[0052] In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 16, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 16. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 15, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 15. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 19, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 20, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 22, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 9. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 134, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 138, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 139. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO:140, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:141.

[0053] In some aspects, this disclosure provides a fusion protein dimer comprising the fusion protein described herein and a second half-life extension domain, wherein the second half-life extension domain does not contain an hIGFBPL1 polypeptide or a fragment thereof. In some embodiments, the first and second half-life extension domains are identical. In some embodiments, the first and second half-life extension domains are different. In some embodiments, the first half-life extension domain is a first Fc domain, and the second half-life extension domain is a second Fc domain. In some embodiments, the first Fc domain contains a club-shaped mutation and the second Fc domain contains a mortar-shaped mutation to dimerize the first and second Fc domains. In some embodiments, the first Fc domain contains a mortar-shaped mutation and the second Fc domain contains a club-shaped mutation to dimerize the first and second Fc domains. In some embodiments, the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NO:4-9, 92, and 93. In some embodiments, the first fusion protein comprises a second hIGFBPL1 polypeptide or a fragment thereof operatively linked to the first half-life extension domain, with or without a linker. In some embodiments, a second hIGFBPL1 peptide or a fragment thereof is operatively linked to the opposite end of the hIGFBPL1 peptide or a fragment thereof. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 22.

[0054] In some aspects, this disclosure provides a fusion protein dimer comprising a first fusion protein and a second fusion protein, wherein the first and second fusion proteins each comprise an amino acid sequence selected from SEQ ID NO:51-86, and wherein the first and second fusion proteins are the same or different.

[0055] In some aspects, this disclosure provides a composition comprising the fusion protein or fusion protein dimer disclosed herein.

[0056] In some aspects, this disclosure provides a method for inhibiting DCC activity in a sample or subject, comprising introducing a fusion protein or fusion protein dimer disclosed herein into the sample or subject. In some embodiments, the fusion protein or fusion protein dimer binds to DCC in cells. In some embodiments, the cells are neurons. In some embodiments, the fusion protein or fusion protein dimer increases mean neurite length, number of neurites, number of neurons, and / or number of neurites per neuron. In some embodiments, the fusion protein increases neurotrophin signaling.

[0057] In some aspects, this disclosure provides a method for inhibiting SORT1 activity in a sample or subject, comprising introducing the disclosed fusion protein or fusion protein dimer into the sample or subject. In some embodiments, the fusion protein or fusion protein dimer binds to SORT1 in cells. In some embodiments, the cells are microglia. In some embodiments, the fusion protein or fusion protein dimer increases the secretion of at least one cytokine by microglia. In some embodiments, the at least one cytokine is selected from IL-10, GM-CSF, MIP-1 α, and any combination thereof. In some embodiments, the at least one cytokine is selected from IL-10, GM-CSF, MIP-1 α, CXCL8 (IL-8), and any combination thereof. In some embodiments, the fusion protein or fusion protein dimer inhibits the production of one or more proteins by microglia. In some embodiments, the one or more proteins comprise one or more inflammatory cytokines. In some embodiments, the one or more inflammatory cytokines comprise IL-6. In some embodiments, the one or more proteins comprise IP-10. In some embodiments, the fusion protein or fusion protein dimer increases the phagocytic activity of microglia. In some embodiments, the cell is a neuron. In some embodiments, the fusion protein or fusion protein dimer increases the average neurite length, the number of neurites, the number of neurons, and / or the number of neurites per neuron. In some embodiments, the fusion protein increases neurotrophic factor signaling.

[0058] In some aspects, this disclosure provides a method for preferentially targeting neurons rather than microglia in a sample or subject, comprising introducing a fusion protein or fusion protein dimer as described herein into the sample or subject.

[0059] In some aspects, this disclosure provides a method for preferentially targeting neurons rather than microglia in a sample or subject, comprising introducing a fusion protein into the sample or subject, the fusion protein comprising a DCC-binding construct and a half-life extension domain.

[0060] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein, fusion protein dimer, or composition described herein. In some embodiments, the retinal degenerative disease is glaucoma, dry age-related macular degeneration (AMD), geographic atrophy, wet AMD, retinitis pigmentosa, diabetic retinopathy, or Stargardt disease.

[0061] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinal disease, comprising administering to the subject a fusion protein, fusion protein dimer, or composition described herein. In some embodiments, the inflammatory retinal disease is retinal vasculitis, uveitis, NAION, or neuromyelitis optica.

[0062] In some embodiments, the fusion protein, fusion protein dimer, or composition is applied to the eye of a subject. In some embodiments, application to the eye includes epidermal application, topical ocular application, or systemic application. In some embodiments, topical ocular application includes periocular, intravitreal, or intraanterior chamber application.

[0063] In some aspects, this disclosure provides a method of treating a subject with a neurodegenerative disease, comprising administering to the subject the fusion protein, fusion protein dimer, or composition described herein. In some embodiments, the fusion protein, fusion protein dimer, or composition is administered parenterally to the subject. In some embodiments, the fusion protein, fusion protein dimer, or composition is administered subcutaneously or intravenously to the subject. In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, frontotemporal dementia, amyotrophic lateral sclerosis, progressive supranuclear palsy, leukoencephalopathy, multiple sclerosis, transverse myelitis, and Schilder's disease. Attached Figure Description

[0064] Figure 1 A schematic diagram showing the combination of human IGBFPL1 protein domains of an exemplary human IGBFPL1 (hIGFBPL1) construct is provided.

[0065] Figure 2A The following schematic diagrams of hIGFBPL1 constructs are provided: PRO802 (divalent IGBBPL1 N-terminus connected to Fc); PRO801 (divalent IGBBPL1 C-terminus connected to Fc); PRO1399 (monovalent IGBBPL1 C-terminus connected to Fc); and PRO1400 (monovalent IGBBPL1 N-terminus connected to Fc).

[0066] Figure 2B and Figure 2C Provided display Figure 2A A diagram showing the binding of the hIGFBPL1 construct to MCF7 cells.

[0067] Figure 3 Images showing the binding of 200 nM PRO801 to cell bodies and synaptic subsets in hIPSC-derived neuronal cultures are provided.

[0068] Figure 4 Provides a display of 200 nM concentration Figure 2A Images of the hIGFBPL1 construct bound to somatic cells, neurites, and synapses.

[0069] Figure 5 Provides a display of 200 nM concentration Figure 2A Images of the binding of the hIGFBPL1 construct to microglia.

[0070] Figure 6A Images showing the combination of PRO802 with iPSC-derived neurons are provided.

[0071] Figures 6B to 6E Provides quantification of neurite length in iPSC-derived neurons after culturing with PRO802 and PRO801. Figure 6B ), number of neural spurs ( Figure 6C ), number of neurons ( Figure 6D ) and the number of neurites per neuron ( Figure 6E The graph was used to determine significance via a post-hoc Dunnett test. p≤0.05, p≤0.01, (p≤0.001). All datasets showed significance using one-way ANOVA. Data were from three fields of view, with each field containing five cultures.

[0072] Figures 7A to 7C A figure showing the effects of PRO801 and PRO802 on cytokine secretion in hIPSC-derived microglia is provided.

[0073] Figure 8 Graphs showing the binding of PRO801 and PRO802 with SORT1 and DCC as determined by Octet are provided.

[0074] Figure 9A and Figure 9B They provided Figure 2A The flow cytometry histograms of the constructs and assay controls are shown.

[0075] Figure 10 The following schematic diagrams of hIGFBPL1 constructs are provided: PRO1571 (tetravalent IGBBPL1 connected to Fc); PRO1572 (trans-divalent IGBBPL1 connected to Fc); and PRO1573 (cis-divalent IGBBPL1 connected to Fc).

[0076] Figures 11A to 11C Provides display PRO801 and Figure 10 The construct in 200 nM ( Figure 11A ) and 20 nM ( Figure 11B Images of the binding of microglia at a concentration of ). Figure 11C The images show binding to neurons at 200 nM or 20 nM. The top image shows the labeling of the IGBBPL1 construct, and the bottom image shows the labeling of microglia based on IBA-1 staining.

[0077] Figure 12 Figures showing the binding of PRO801 via PNGase glycosylation (013) or deglycosylation (012) to different concentrations of BeWo cells are provided.

[0078] Figure 13A Images showing the binding of PRO801 with glycosylated (013) or deglycosylated (012) neurons to 200 nM or 20 nM neurons are provided.

[0079] Figure 13B Images showing the binding of PRO801 with 20 nM microglia, either glycosylated (013) or deglycosylated (012).

[0080] Figure 14 A diagram showing cell binding to BeWo cells (epithelial cells) with the indicated glycosylation mutation of the IGFFBPL1-Fc fusion protein is provided.

[0081] Figure 15 A graph showing the binding of recombinant mouse IGBFPL1 (PRO806) to human and mouse SORT1 and DCC via biolayer interferometry (BLI) is provided.

[0082] Figure 16 A diagram showing the binding of the IGFFBPL1-Fc fusion protein to microglia is provided.

[0083] Figure 17 A graph showing the correlation between the IGBBPL1-Fc fusion protein and microglia and SORT1 is provided.

[0084] Figure 18 A figure showing the effect of the IGFFBPL1-Fc fusion protein on microglia phagocytosis is provided.

[0085] Figures 19A to 19B Provided display of IL-6 in microglia treated with LPS ( Figure 19A ) and IP-10 ( Figure 19B The expression of the protein was inhibited by the indicated concentration of the IGFBPL1-Fc fusion protein PRO1643.

[0086] Figures 20A to 20B Provided display of IGBBPL1-Fc fusion protein with 20 nM ( Figure 20A ) or 200 nM ( Figure 20B A diagram showing the concentration of neurons bound together.

[0087] Figure 21 A graph showing AKT activation in neurons at the indicated concentrations of the IGFBPL1-Fc fusion protein PRO1643 is provided.

[0088] Figure 22 A graph showing neuronal survival in cultures treated with the indicated concentration of IGFFBPL1-Fc fused with PRO1643 is provided. Detailed Implementation

[0089] IGBFPL1 has previously been described as binding to neurons and microglia (see, for example, U.S. Patent No. 10,842,849 and U.S. Publication No. 2022-01431319, each incorporated herein by reference). However, all binding partners of IGBFPL1 are unknown. This disclosure is based at least in part on the finding that IGBFPL1 binds not to IGF1R as previously predicted, but to sorting protein 1 (SORT1) in addition to the neuroguide factor 1 receptor (DCC). Specifically, as demonstrated herein, fusion proteins presenting the N-terminus of IGBFPL1 (i.e., fused to the C-terminus) have been identified as binding only to DCC, while fusion proteins presenting the C-terminus of IGBFPL1 (i.e., fused to the N-terminus) have been identified as binding to both DCC and SORT1. Furthermore, it has been determined that the N-terminal domain of IGBFPL1 is essential for DCC binding, while the Ig-like domain is essential for SORT1 binding. SORT1 is highly expressed in the brain and eye, particularly on microglia. SORT1 is a major contributor to the clearance of granulin precursor (PGRN), blocking pro-inflammatory signaling and inducing microglia to engage in inflammatory resolution. Without being bound by theory, IGBBPL1 provides anti-inflammatory and inflammatory resolution by inhibiting SORT1. Furthermore, SORT1 acts as an ApoE receptor, a known activator of microglia. Therefore, based on the data presented in this paper, it is considered that IGBBPL1 competes with ApoE and this is another mechanism by which IGBBPL1 provides anti-inflammatory activity. DCC is highly expressed in the brain, with the highest expression in neurons, and is also highly expressed in neurons in the eye. DCC is a receptor for neurotrophic factors, and mutations in DCC are associated with depression, psychiatric disorders, and Parkinson's disease. Neuronal growth has been previously reported when neurotrophic factors are applied exogenously. Therefore, without being bound by theory, the ability of IGBBPL1 to promote neuronal survival is thought to be based on its binding to DCC, similar to its endogenous ligand, neurotrophic factors.

[0090] In some aspects, this disclosure provides a component for binding to DCC. In some embodiments, the component for binding to DCC is the IGBBPL1 polypeptide or IGBBPL1 fragment described herein. In some embodiments, the component for binding to DCC is a fusion protein described herein.

[0091] In some aspects, this disclosure provides a component for binding to SORT1. In some embodiments, the component for binding to SORT1 is the IGBBPL1 polypeptide or IGBBPL1 fragment described herein. In some embodiments, the component for binding to SORT1 is a fusion protein described herein.

[0092] In some aspects, this disclosure provides a component for binding to DCC and SORT1. In some embodiments, the component for binding to DCC and SORT1 is the IGBBPL1 polypeptide or IGBBPL1 fragment described herein. In some embodiments, the component for binding to DCC and SORT1 is a fusion protein described herein.

[0093] This article provides IGFFBPL1 fragments containing one or more domains of wild-type IGFFBPL1 protein (e.g., human wild-type IGFPBL1 protein) and fusion proteins containing one or more domains of wild-type IGFFBPL1 protein (e.g., human wild-type IGFPBL1 protein) and a half-life extension domain. Methods for inhibiting SORT1 and / or DCC, as well as methods for preferentially targeting neurons and microglia, are also provided.

[0094] definition

[0095] Unless otherwise stated, the terms used in the claims and description are defined as follows.

[0096] As used herein, “about” will be understood by a person skilled in the art and will vary to some extent depending on the context in which it is used. If the use of the term is unclear to a person skilled in the art in the context in which it is used, then “about” will mean at most 10% plus or minus a particular value.

[0097] "Amino acids" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid simulants that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that have been modified, such as hydroxyproline, g-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure.

[0098] Amino acid analogs are naturally occurring amino acids, consisting of carbon atoms bound to hydrogen, carboxyl, amino, and R groups, such as homoserine, ortholeucine, methionine sulfoxide, and methionine methylsulfonium. These analogs have modified R groups (e.g., ortholeucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimics are chemical compounds that have a structure different from the general chemical structure of amino acids but function in a manner similar to naturally occurring amino acids.

[0099] Amino acids in this article can be identified by their commonly known three-letter symbols or by IUPAC-IUB Biochemistry

[0100] The nomenclature committee recommends using single-letter symbols to represent nucleotides. Similarly, nucleotides can be identified by their generally accepted single-letter codes.

[0101] "Amino acid substitution" refers to replacing at least one existing amino acid residue in a predetermined amino acid sequence (the amino acid sequence of the starting polypeptide) with a second, different "substitute" amino acid residue. "Amino acid insertion" refers to incorporating at least one additional amino acid into a predetermined amino acid sequence. While insertions typically consist of the insertion of one or two amino acid residues, larger "peptide insertions" can also be made, for example, inserting about three to about five or even up to about ten, fifteen, or twenty amino acid residues. As disclosed above, the inserted residues can be naturally occurring or non-naturally occurring. "Amino acid deletion" refers to removing at least one amino acid residue from a predetermined amino acid sequence.

[0102] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably in this document and refer to polymers of amino acid residues. This term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to both naturally occurring and non-naturally occurring amino acid polymers.

[0103] "Nucleic acid" refers to deoxyribonucleotides or ribonucleotides in single-stranded or double-stranded form, and polymers thereof. Unless otherwise specified, the term covers nucleic acids containing known analogs of naturally occurring nucleotides, having similar binding properties to a reference nucleic acid, and being metabolized in a manner similar to that of naturally occurring nucleotides. Unless otherwise stated, a specific nucleic acid sequence is also implicitly included.

[0104] This includes variants with conserved modifications (e.g., degenerate codon substitutions) and complementary sequences, as well as explicitly indicated sequences. Specifically, degenerate codon substitutions can be achieved by mixing bases and / or...

[0105] The sequence is achieved through substitution of deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., Biol. Chem. 260:2605-2608, 1985; and Cassol et al., 1992; Rossolini et al., Mol. Cell. Probes 8:91-98, 1994). For arginine and leucine, modifications at the second base can also be conserved. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene.

[0106] The polynucleotides used in this article can be any polynucleotide or

[0107] Polynucleotides are composed of unmodified RNA or DNA, or modified RNA or DNA. For example, polynucleotides can be composed of single-stranded and double-stranded DNA, DNA as a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA as a mixture of single-stranded and double-stranded regions, or hybrid molecules containing DNA and RNA that may be single-stranded or more commonly double-stranded, or a mixture of single-stranded and double-stranded regions. Furthermore, polynucleotides can be composed of triple-stranded regions containing RNA or DNA, or both RNA and DNA. Polynucleotides may also contain one or more modified bases or the DNA or RNA backbone modified for stability or other reasons. "Modified" bases include, for example, triphenylmethylated bases and uncommon bases (such as inosine). DNA and RNA can be modified in various ways; therefore, "polynucleotide" includes forms of chemical, enzymatic, or metabolic modifications. Polynucleotides can be synthetic or isolated nucleic acid polymers comprising multiple nucleotide subunits.

[0108] As used herein, the terms “connected,” “combined,” “coupled,” “fused,” or “fusion” are used interchangeably when referring to the connection of two or more elements or components or domains together by any means including chemical combination or recombination. Methods of chemical combination (e.g., using heterobifunctional crosslinking agents) are known in the art.

[0109] The term "derived from" a peptide, polypeptide, or amino acid sequence refers to the source of the polypeptide. Preferably, the peptide, polypeptide, or amino acid sequence derived from a specific sequence has an amino acid sequence substantially identical to that sequence or a portion thereof, wherein the portion consists of at least 5-15 amino acids, at least 10-20 amino acids, at least 20-30 amino acids, at least 30-50 amino acids, or can be otherwise identified by those skilled in the art as having its origin in that sequence. A polypeptide derived from another peptide may have one or more mutations relative to the starting polypeptide, for example, one or more amino acid residues have been substituted by another amino acid residue or have one or more amino acid residues inserted or deleted.

[0110] Peptides or polypeptides may contain amino acid sequences that are not naturally occurring. Such variants necessarily have less than 100% sequence identity or similarity to the starting molecule. In some embodiments, the variant will have, for example, an amino acid sequence identity or similarity of about 75% to less than 100% with the amino acid sequence of the starting polypeptide in length of the variant molecule, more preferably about 80% to less than 100%, more preferably about 85% to less than 100%, more preferably about 90% to less than 100% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%), and most preferably about 95% to less than 100% amino acid sequence identity or similarity.

[0111] In some embodiments, there is an amino acid difference between the starting polypeptide sequence and the sequence derived therefrom. The identity or similarity of this sequence is defined herein as the percentage of amino acid residues in the candidate sequence that are identical (i.e., same residues) to the starting amino acid residues after sequence alignment and, where necessary, the introduction of vacancies to achieve the maximum percentage of sequence identity.

[0112] In some embodiments, the peptide or polypeptide is encoded by a nucleotide sequence. The nucleotide sequences of the present invention can be used in many applications, including: cloning, gene therapy, protein expression and purification, mutation introduction, DNA vaccination of desired hosts, antibody production for, for example, passive immunization, PCR, primer and probe production, etc.

[0113] As used herein, the term “operably connected” or “operably coupled” refers to a juxtaposition in which the described components are in a relationship that allows them to function in their intended manner.

[0114] The term "in vivo" refers to processes that occur within a living organism.

[0115] As used herein, the terms “mammal” or “subject” or “patient” include humans and non-humans, and include, but are not limited to, humans, non-human primates, canines, felines, rodents, bovines, equines, and pigs. The terms “mammal,” “subject,” and “patient” are used interchangeably herein. The terms “mammal,” “subject,” and “patient” refer to any individual who is the target of treatment using the compositions of this disclosure. A subject may be a vertebrate, such as a mammal. A subject may be human. A subject may be symptomatic or asymptomatic. The term does not indicate a specific age or sex. It is intended to cover adult and neonatal subjects, whether male or female. Subjects may include control subjects or test subjects.

[0116] In the context of two or more nucleic acid or polypeptide sequences, the term "percentage of identity" refers to two or more sequences or subsequences that have a specified percentage of identical nucleotide or amino acid residues when comparing and aligning for maximum correspondence, whether measured by visual inspection or by using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to those skilled in the art). Depending on the application, "percentage identity" can exist in regions of the sequences being compared, such as in functional domains, or across the full length of the two sequences being compared. For sequence comparison, typically one sequence serves as a reference sequence to be compared with the test sequence. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, and if necessary, the coordinates of the subsequences are specified, along with the sequence algorithm program parameters. The sequence comparison algorithm then calculates the percentage of sequence identity of the test sequence relative to the reference sequence based on the specified program parameters.

[0117] The optimal alignment of sequences for comparison can be achieved, for example, through the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), and the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA85:2444 (1988), through the computerized implementation of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr.). (Madison, Wis.) or by visual inspection (see generally Ausubel et al., below).

[0118] One example of an algorithm suitable for determining the percentage of sequence identity and the percentage of sequence similarity is the BLAST algorithm, described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analysis is publicly available from the website of the National Center for Biotechnology Information.

[0119] As used herein, the term "gly-ser linker" refers to a peptide composed of glycine and serine residues. An exemplary gly-ser peptide linker comprises the amino acid sequence Ser(Gly4Ser)n. In some embodiments, n=1. In some embodiments, n=2. In some embodiments, n=3, i.e., Ser(Gly4Ser)3. In some embodiments, n=4, i.e., Ser(Gly4Ser)4. In some embodiments, n=5. In some embodiments, n=6. In some embodiments, n=7. In some embodiments, n=8. In some embodiments, n=9. In some embodiments, n=10. Another exemplary gly-ser peptide linker comprises the amino acid sequence (Gly4Ser)n. In some embodiments, n=1. In some embodiments, n=2. In some embodiments, n=3. In some embodiments, n=4. In some embodiments, n=5. In some embodiments, n=6. Another exemplary gly-ser peptide linker comprises the amino acid sequence (Gly3Ser)n. In some implementations, n=1. In some implementations, n=2. In some implementations, n=3. In some implementations, n=4. In some implementations, n=5. In some implementations, n=6.

[0120] As used herein, the terms “Fc region,” “Fc chain,” and “Fc domain” are used interchangeably and each refers to a portion of a native immunoglobulin formed by the corresponding Fc domain (or Fc portion) of its two heavy chains. As used herein, the term “Fc domain” refers to a portion of a single immunoglobulin (Ig) heavy chain in which the Fc domain does not contain an Fv domain. Therefore, the Fc domain may also be referred to as “Ig” or “IgG.” In some embodiments, the Fc domain begins in a hinge region immediately upstream of the papain cleavage site and terminates at the C-terminus of the antibody. Thus, a complete Fc domain contains at least a hinge domain, a CH2 domain, and a CH3 domain. In some embodiments, the Fc domain contains at least one of the following: a hinge (e.g., an upper hinge region, a middle hinge region, and / or a lower hinge region) domain, a CH2 domain, a CH3 domain, a CH4 domain, or a variant, portion, or fragment thereof. In some embodiments, the Fc domain comprises a complete Fc domain (i.e., a hinge domain, a CH2 domain, and a CH3 domain). In some embodiments, the Fc domain comprises a hinge domain (or a portion thereof) fused to a CH3 domain (or a portion thereof). In some embodiments, the Fc domain comprises a CH2 domain (or a portion thereof) fused to a CH3 domain (or a portion thereof). In some embodiments, the Fc domain consists of a CH3 domain (or a portion thereof). In some embodiments, the Fc domain consists of a hinge domain (or a portion thereof) and a CH3 domain (or a portion thereof). In some embodiments, the Fc domain consists of a CH2 domain (or a portion thereof) and a CH3 domain (or a portion thereof). In some embodiments, the Fc domain consists of a hinge domain (or a portion thereof) and a CH2 domain (or a portion thereof). In some embodiments, the Fc domain lacks at least a portion of the CH2 domain (e.g., all or part of the CH2 domain). The Fc domain as used herein generally refers to a polypeptide containing all or part of the Fc domain of an immunoglobulin heavy chain. This includes, but is not limited to, peptides containing the entire CH1, hinge, CH2, and / or CH3 domains, as well as fragments of such peptides containing only, for example, hinge, CH2, and CH3 domains. In some embodiments, the Fc domain is derived from any species and / or any subtype of immunoglobulin, including but not limited to human IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibodies. The human IgG1 constant region is shown in Uniprot P01857 and Table 12 (i.e., SEQ ID NO:172). The Fc domain of human IgG1 is shown in Table 12 (i.e., SEQ ID NO:173). The Fc domain encompasses both native Fc and Fc variant molecules. As with Fc variants and native Fc, the term Fc domain includes molecules in monomeric or multimeric form, whether produced from whole antibody digestion or by other means.The amino acid residue numbering of the Fc domain is assigned according to Kabat's definition. See, for example, Sequences of Proteins of Immunological Interest (Table of Contents, Introduction and Constant Region Sequences sections), 5th edition, Bethesda, MD: NIH Vol. 1: 647-723 (1991); Kabat et al., "Introduction" Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services, NIH, 5th edition, Bethesda, MD Vol. 1: xiii-xcvi (1991); Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987); Chothia et al., Nature 342: 878-883 (1989), each of which is incorporated herein by reference for all purposes.

[0121] As described herein, those skilled in the art will understand that any Fc domain can be modified such that its amino acid sequence differs from the native Fc domain of naturally occurring immunoglobulin molecules. In some embodiments, the Fc domain has reduced effector function (e.g., FcγR binding).

[0122] In some embodiments, the Fc domain is derived from different immunoglobulin molecules. For example, the Fc domain may include a CH2 and / or CH3 domain derived from an IgG1 molecule and a hinge region derived from an IgG3 molecule. In another example, the Fc domain may include a chimeric hinge region partially derived from an IgG1 molecule and partially derived from an IgG3 molecule. In yet another example, the Fc domain may include a chimeric hinge partially derived from an IgG1 molecule and partially derived from an IgG4 molecule.

[0123] As used herein, the term "fusion protein" refers to a recombinant protein prepared by fusing the multispecific variable regions and polymers (e.g., serum albumin) described herein.

[0124] As used in the context of sequences herein, the term "identity" refers to the relationship between two or more sequences, as determined by sequence comparison. The term "identity" also implies the degree of sequence relevance between peptides, as determined by the number of matches between strings of two or more amino acid residues. The percentage of "identity" between two sequences is a function of the number of common positions shared by the sequences (i.e., the percentage of identity equals the number of common positions / total number of positions × 100), taking into account the number of vacancies and the length of each vacancy, which needs to be incorporated for optimal alignment of the two sequences. Sequence comparison and determination of the percentage of identity between two sequences can be accomplished using mathematical algorithms. For sequence comparison, typically one sequence serves as a reference sequence to be compared with the test sequence. When using a sequence comparison algorithm, the test and reference sequences are input into the computer, and subsequence coordinates are specified if necessary, along with the sequence algorithm program parameters. Default program parameters can be used, or alternative parameters can be specified. The sequence comparison algorithm then calculates the percentage of sequence identity of the test sequence relative to the reference sequence based on the program parameters. Alternatively or concurrently, the amino acid sequences disclosed herein can be further used as “query sequences” to search public databases for, for example, to identify relevant sequences. Such searches can be performed using the BLAST program of Altschul et al. (J. Mol. Biol. 215:403-10, 1990).

[0125] As used herein, the term "conservative substitution" refers to substitution with another amino acid residue having properties similar to those of the original amino acid residue. For example, lysine, arginine, and histidine have similar properties because they have basic side chains, and aspartic acid and glutamic acid have similar properties because they have acidic side chains. Furthermore, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan have similar properties because they have uncharged polar side chains, and alanine, valine, leucine, threonine, isoleucine, proline, phenylalanine, and methionine have similar properties because they have nonpolar side chains. Additionally, tyrosine, phenylalanine, tryptophan, and histidine have similar properties because they have aromatic side chains. Therefore, it will be apparent to those skilled in the art that even when substitution occurs with amino acid residues in the group exhibiting similar properties as described above, a significant change in the properties of the antibody or antigen-binding fragment may not be observed.

[0126] As used herein, “half-life” refers to the time taken for the serum or plasma concentration of a polypeptide to decrease by 50% in vivo, for example, due to degradation and / or clearance or chelation by natural mechanisms. The extended PKIGFBPL1 suitable for the methods disclosed herein is stable in vivo, and its half-life is increased by, for example, fusion with the Fc region, fusion with serum albumin (e.g., HSA or MSA), by PEGylation, or by binding to serum albumin molecules (e.g., human serum albumin) resistant to degradation and / or clearance or chelation. Half-life can be determined in any manner known per se, such as by pharmacokinetic analysis. Suitable techniques will be apparent to those skilled in the art and may typically involve, for example, the following steps: appropriately administering a suitable dose of the amino acid sequence or compound to a subject; periodically collecting blood samples or other samples from said subject; determining the level or concentration of the amino acid sequence or compound in said blood sample; and calculating, based on the data thus obtained (in a graph), the time until the level or concentration of the amino acid sequence or compound decreases by 50% compared to the initial level after administration. Further details are provided in, for example, standard manuals such as Kenneth, A. et al., Chemical Stability of Pharmaceuticals: A Handbook for Pharmacists and Peters et al., Pharmacokinetic Analysis: A Practical Approach (1996). See also Gibaldi, M. et al., Pharmacokinetics, 2nd Revision, Marcel Dekker (1982).

[0127] IGFBPL1 peptide and fusion protein

[0128] In some embodiments, this disclosure provides a fusion protein comprising a human IGBBPL1 (hIGFBPL1) polypeptide or a fragment thereof and a half-life extension domain. The hIGFBPL1 polypeptide or a fragment thereof is operatively linked to the half-life extension domain with or without a linker.

[0129] In some embodiments, this disclosure provides an IGBBPL1 fragment of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment is a fragment of hIGFBPL1. In some embodiments, the IGBBPL1 fragment is a fragment of mouse IGBBPL1 (mIGFBPL1). In some embodiments, the IGBBPL1 fragment comprises both a fragment of mIGFBPL1 and a fragment of hIGFBPL1.

[0130] In some embodiments, this disclosure provides an IGBBPL1 polypeptide comprising fragments of hIGFBPL1 and mIGFBPL1.

[0131] IGFBPL1 peptide and fragments

[0132] Insulin-like growth factor binding protein-like 1 (IGFBPL1) is a soluble protein having multiple domains. Specifically, wild-type IGFFBPL1 includes a thumb domain, an N-terminal domain, a Kazal domain, an Ig-like domain, and a C-terminal domain. In some embodiments, the IGFFBPL1 peptide suitable for use in the fusion protein described herein comprises any one of the domains present in wild-type IGFFBPL1. In some embodiments, the IGFFBPL1 peptide suitable for use in the fusion protein described herein comprises any combination of the domains present in wild-type IGFFBPL1.

[0133] Those skilled in the art will be able to identify the domains of the selected IGFFBPL1 peptide.

[0134] In some embodiments, the IGFPL1 peptide and fragment described herein are constructs for binding DCC. In some embodiments, the IGFPL1 peptide and fragment described herein are constructs for binding SORT1. In some embodiments, the IGFPL1 peptide and fragment described herein are constructs for binding both DCC and SORT1. Methods for determining molecular binding are known to those skilled in the art.

[0135] In some embodiments, the thumb domain comprises the amino acid sequence shown in SEQ ID NO: 25. In some embodiments, the thumb domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the same amino acid sequence as SEQ ID NO: 25. In some embodiments, the thumb domain comprises an amino acid sequence having 8-11 amino acid residues as in SEQ ID NO: 25. In some embodiments, the thumb domain comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the thumb domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 25. In some embodiments, the thumb domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 25.

[0136] In some embodiments, the N-terminal domain comprises the amino acid sequence shown in SEQ ID NO: 2. In some embodiments, the N-terminal domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the same amino acid sequence as SEQ ID NO: 2. In some embodiments, the N-terminal domain comprises an amino acid sequence having 60-70 amino acid residues as in SEQ ID NO: 2. In some embodiments, the N-terminus comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the N-terminus comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 2. In some embodiments, the N-terminus comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 2.

[0137] In some embodiments, the Kazal domain comprises the amino acid sequence shown in SEQ ID NO: 27. In some embodiments, the Kazal domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the same amino acid sequence as SEQ ID NO: 27. In some embodiments, the Kazal domain comprises an amino acid sequence having 32-42 amino acid residues as in SEQ ID NO: 27. In some embodiments, the Kazal domain comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the Kazal domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 27. In some embodiments, the Kazal domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 27.

[0138] In some embodiments, the Ig-like domain comprises the amino acid sequence shown in SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the same amino acid sequence as SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises an amino acid sequence having 98-108 amino acid residues as in SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the Ig-like domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 26.

[0139] In some embodiments, the C-terminal domain comprises the amino acid sequence shown in SEQ ID NO: 3. In some embodiments, the C-terminal domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the same amino acid sequence as SEQ ID NO: 3. In some embodiments, the C-terminal domain comprises an amino acid sequence having 10-16 amino acid residues of SEQ ID NO: 3. In some embodiments, the C-terminus comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the C-terminus comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 3. In some embodiments, the C-terminus comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 3.

[0140] In some embodiments, the IGBBPL1 fragment includes the N-terminal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes the C-terminal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes the thumb domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes the Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes the Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide.

[0141] In some embodiments, the IGBBPL1 fragment comprises one to four IGBBPL1 domains selected from the N-terminal, C-terminal, thumb, Kazal, and Ig-like domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment comprises two IGBBPL1 domains selected from the N-terminal, C-terminal, thumb, Kazal, and Ig-like domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment comprises three IGBBPL1 domains selected from the N-terminal, C-terminal, thumb, Kazal, and Ig-like domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment comprises four IGBBPL1 domains selected from the N-terminal, C-terminal, thumb, Kazal, and Ig-like domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment comprises no more than four of the following: the N-terminal domain, C-terminal domain, thumb domain, Kazal domain, and Ig-like domain of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment comprises more than one copy of the IGBBPL1 domains described herein.

[0142] In some embodiments, the IGBBPL1 fragment includes an N-terminal domain and a thumb domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain and a C-terminal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a C-terminal domain and a thumb domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a C-terminal domain and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a C-terminal domain and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a thumb domain and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a thumb domain and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a Kazal domain and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide.

[0143] In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a Kazal domain, and a thumb domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, and a thumb domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a thumb domain, and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, an Ig-like domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a C-terminal domain, a thumb domain, and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a C-terminal domain, a thumb domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes a Kazal domain, a thumb domain, and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some implementations, the IGBBPL1 fragment includes the C-terminal domain, Ig-like domain, and Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not contain other domains of the IGBBPL1 peptide.

[0144] In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, a thumb domain, and an Ig-like domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, a thumb domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a C-terminal domain, an Ig-like domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some embodiments, the IGBBPL1 fragment includes an N-terminal domain, a thumb domain, an Ig-like domain, and a Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide. In some implementations, the IGBBPL1 fragment includes the C-terminal domain, thumb domain, Ig-like domain, and Kazal domain of the IGBBPL1 peptide, provided that the IGBBPL1 fragment does not include other domains of the IGBBPL1 peptide.

[0145] In some embodiments, the IGBBPL1 fragment comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID Nos: 2-3, 25-27, 31-35, 37-50, 158, and 160-179. In some embodiments, the IGBBPL1 fragment comprises an amino acid sequence selected from SEQ ID No: 2-3, 25-27, 31-35, 37-50, 158, and 160-179. In some embodiments, the IGBBPL1 fragment consists of an amino acid sequence selected from SEQ ID No: 2-3, 25-27, 31-35, 37-50, 158, and 160-179. In some embodiments, the IGBBPL1 fragment consists of an amino acid sequence selected from SEQ ID No: 2-3, 25-27, 31-35, 37-50, 158, and 160-179.

[0146] In some embodiments, the IGBBPL1 polypeptide provided herein is a chimeric IGBBPL1 polypeptide comprising at least one IGBBPL1 domain of a human IGBBPL1 polypeptide and at least one IGBBPL1 domain of a non-human IGBBPL1 polypeptide. In some embodiments, the non-human IGBBPL1 polypeptide is a mouse IGBBPL1 polypeptide. In some embodiments, the chimeric IGBBPL1 polypeptide comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 151-157. In some embodiments, the chimeric IGBBPL1 polypeptide consists of an amino acid sequence selected from SEQ ID No: 151-157.

[0147] In some embodiments, the fusion protein described herein comprises the IGBBPL1 peptide or IGBBPL1 fragment described herein.

[0148] In some embodiments, the fusion protein includes the N-terminal domain of the IGBBPL1 peptide. In some embodiments, the fusion protein includes the N-terminal domain of the IGBBPL1 peptide but does not include the C-terminal domain of the IGBBPL1 peptide. In some embodiments, the fusion protein includes the C-terminal domain of the IGBBPL1 peptide but does not include the N-terminal domain of the IGBBPL1 peptide.

[0149] In some embodiments, the fusion protein includes the N-terminal domain of the human IGBFPL1 peptide. In some embodiments, the fusion protein includes the N-terminal domain of the human IGBFPL1 peptide but does not include the C-terminal domain of the human IGBFPL1 peptide. In some embodiments, the fusion protein includes the C-terminal domain of the human IGBFPL1 peptide but does not include the N-terminal domain of the human IGBFPL1 peptide.

[0150] In some embodiments, the fusion protein includes the N-terminal domain of the human wild-type IGBBPL1 peptide. In some embodiments, the fusion protein includes the N-terminal domain of the human wild-type IGBBPL1 peptide but does not include the C-terminal domain of the human wild-type IGBBPL1 peptide. In some embodiments, the fusion protein includes the C-terminal domain of the human wild-type IGBBPL1 peptide but does not include the N-terminal domain of the human wild-type IGBBPL1 peptide.

[0151] In some embodiments, the fusion protein includes the N-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 2. In some embodiments, the fusion protein includes the N-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 2, but does not include the C-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 3. In some embodiments, the fusion protein includes the C-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 3. In some embodiments, the fusion protein includes the C-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 3, but does not include the N-terminal domain of the human IGBBPL1 peptide as shown in SEQ ID NO: 2.

[0152] In some embodiments, the fusion protein includes a domain of the SORT1-binding IGBBPL1 peptide. In some embodiments, the SORT1-binding IGBBPL1 peptide domain is an Ig-like domain. In some embodiments, the SORT1-binding IGBBPL1 peptide domain is an Ig-like domain and a Kazal domain. In some embodiments, the SORT1-binding IGBBPL1 peptide domain is an Ig-like domain, a Kazal domain, and a C-terminal domain. In some embodiments, the fusion protein includes a domain of the SORT1-binding IGBBPL1 peptide that is not bound to DCC.

[0153] In some embodiments, the fusion protein includes a domain of the IGBBPL1 peptide that binds to DCC. In some embodiments, the domain of the IGBBPL1 peptide that binds to DCC is an N-terminal domain. In some embodiments, the domains of the IGBBPL1 peptide that binds to DCC are an N-terminal domain and a thumb domain. In some embodiments, the domains of the IGBBPL1 peptide that binds to DCC are an N-terminal domain, a thumb domain, and a Kazal domain. In some embodiments, the fusion protein includes a domain of the IGBBPL1 peptide that binds to DCC but does not bind to SORT1.

[0154] In some implementations, the fusion protein includes a domain of the IGBBPL1 peptide that binds to DCC and SORT1.

[0155] In some embodiments, the fusion protein comprises a fragment of the IGBBPL1 peptide. In some embodiments, the fragment comprises one or more domains of the IGBBPL1 peptide. In some embodiments, the fragment does not comprise all domains of the IGBBPL1 peptide. In some embodiments, the fragment comprises a portion of one or more domains of the IGBBPL1 peptide. In some embodiments, the fragment comprises a thumb domain of the IGBBPL1 peptide. In some embodiments, the fragment comprises a thumb domain of the human IGBBPL1 peptide. In some embodiments, the fragment comprises a thumb domain containing the amino acid sequence of SEQ ID NO: 25. In some embodiments, the fragment comprises an N-terminal domain of the IGBBPL1 peptide. In some embodiments, the fragment comprises an N-terminal domain of the human IGBBPL1 peptide. In some embodiments, the fragment comprises an N-terminal domain containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the fragment comprises a Kazal domain of the IGBBPL1 peptide. In some embodiments, the fragment comprises a Kazal domain of the human IGBBPL1 peptide. In some embodiments, the fragment comprises a Kazal domain containing the amino acid sequence of SEQ ID NO: 27. In some embodiments, the fragment comprises an Ig-like domain of the IGBBPL1 peptide. In some embodiments, the fragment comprises an Ig-like domain of the human IGBBPL1 peptide. In some embodiments, the fragment comprises an Ig-like domain containing the amino acid sequence of SEQ ID NO: 26. In some embodiments, the fragment comprises a C-terminal domain of the IGBBPL1 peptide. In some embodiments, the fragment comprises a C-terminal domain of the human IGBBPL1 peptide. In some embodiments, the fragment comprises a C-terminal domain containing the amino acid sequence of SEQ ID NO: 3.

[0156] In some embodiments, the fusion protein comprises a fragment of the hIGFBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues P36-M278 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues P36-A109 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues L26-E152 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues P36-E152 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues V107-E152 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues C108-E152 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues V107-M278 of the IGBBPL1 polypeptide. In some embodiments, the fragment, by linear numbering, comprises amino acid residues C108-M278 of the IGBBPL1 polypeptide. In some embodiments, the fragment contains amino acid residues V107-S264 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues C108-S264 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues E152-M278 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues E152-S264 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues L26-Q174 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues L26-G104 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues L26-G112 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues L26-G119 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues L26-A95 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues K37-G104 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues K37-G112 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues K37-G119 of the IGBBPL1 peptide by linear numbering. In some embodiments, the fragment contains amino acid residues K37-A95 of the IGBBPL1 peptide by linear numbering.In some embodiments, the fragment contains amino acid residues C151A-T259 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues C151A-R277 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues A154-T259 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues A154-R277 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues G71-T259 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues G71-R277 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues A96-T259 of the IGBBPL1 peptide, numbered sequentially. In some embodiments, the fragment contains amino acid residues K37-E152 of the IGBBPL1 peptide, numbered sequentially. In some implementations, the fragment contains amino acid residues E152-R277 of the IGBBPL1 polypeptide, using linear numbering.

[0157] In some embodiments, the fragment comprises amino acid residues P36-M278 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues P36-A109 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-E152 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues P36-E152 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues V107-E152 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues C108-E152 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues V107-M278 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues C108-M278 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues V107-S264 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues C108-S264 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues E152-M278 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues E152-S264 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-Q174 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-G104 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-G112 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-G119 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues L26-A95 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues K37-G104 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues K37-G112 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues K37-G119 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues K37-A95 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues C151A-T259 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues C151A-R277 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues A154-T259 of SEQ ID NO: 25.In some embodiments, the fragment comprises amino acid residues A154-R277 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues G71-T259 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues G71-R277 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues A96-T259 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues K37-E152 of SEQ ID NO: 25. In some embodiments, the fragment comprises amino acid residues E152-R277 of SEQ ID NO: 25.

[0158] In some embodiments, the fragment comprises at least two domains of IGBBPL1 operably linked via a linker. For example, in some embodiments, the fragment comprises an N-terminal domain operably linked to a fragment comprising an Ig-like domain and a C-terminal domain. In some embodiments, the fragment comprising at least two domains of IGBBPL1 operably linked via a linker comprises an amino acid sequence selected from SEQ ID No: 175-177.

[0159] In some embodiments, the fragment contains the mutation C108S. In some embodiments, the fragment contains the mutation V107X. In some embodiments, the fragment contains the mutation V107T. In some embodiments, the fragment contains the mutation C108S. In some embodiments, the fragment contains the mutation C151A.

[0160] In some embodiments, the fragment comprises an amino acid sequence selected from SEQ ID NO: 37-50. In some embodiments, the fragment comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with any of the amino acid sequences shown in SEQ ID No: 37-50.

[0161] In some embodiments, the fragment comprises an amino acid sequence selected from SEQ ID NO:158 and 160-179. In some embodiments, the fragment comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with any of the amino acid sequences shown in SEQ ID No:158 and 160-179.

[0162] In some embodiments, the fragment comprises an amino acid sequence selected from SEQ ID NO:37-50, 158, and 160-179. In some embodiments, the fragment comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with any of the amino acid sequences shown in SEQ ID No:37-50, 158, and 160-179.

[0163] In some embodiments, the fusion protein comprises the hIGFBPL1 polypeptide, which contains the amino acid sequence of SEQ ID NO: 10. In some embodiments, the fusion protein comprises the hIGFBPL1 polypeptide, which contains at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the sequence shown in SEQ ID NO: 10. In some embodiments, the fusion protein hIGFBPL1 polypeptide comprises the sequence shown in SEQ ID NO: 10 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0164] In some embodiments, the fusion protein comprises the hIGFBPL1 polypeptide, which contains the amino acid sequence of SEQ ID NO: 11. In some embodiments, the fusion protein comprises the hIGFBPL1 polypeptide, which contains at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the sequence shown in SEQ ID NO: 11. In some embodiments, the fusion protein comprises the hIGFBPL1 polypeptide, which contains the sequence shown in SEQ ID NO: 11 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0165] In some embodiments, the IGBBPL1 peptide or a fragment thereof contains a mutation that alters glycosylation. In some embodiments, the glycosylation mutation is at position N166 of the hIGFBPL1 peptide. In some embodiments, the glycosylation mutation is N166D. In some embodiments, the IGBBPL1 peptide with the N166D mutation is shown in SEQ ID NO: 87. In some embodiments, the glycosylation mutation is N166Q. In some embodiments, the IGBBPL1 peptide with the N166Q mutation is shown in SEQ ID NO: 88. In some embodiments, the glycosylation mutation is at position S268 of the hIGFBPL1 peptide. In some embodiments, the glycosylation mutation is S268A. In some embodiments, the IGBBPL1 peptide with the S268A mutation is shown in SEQ ID NO: 89. In some embodiments, the IGBBPL1 peptide or a fragment thereof contains glycosylation mutations at positions N166 and S268. In some embodiments, the IGBBPL1 peptide or a fragment thereof contains glycosylation mutations N166D and S268A. In some embodiments, the IGBBPL1 peptide having N166D and S268A mutations is shown in SEQ ID NO: 90. In some embodiments, the IGBBPL1 peptide or a fragment thereof comprises the glycosylation mutations N166Q and S268A. In some embodiments, the IGBBPL1 peptide having N166Q and S268A mutations is shown in SEQ ID NO: 91.

[0166] In some embodiments, the IGBBPL1 fragment described herein does not contain a half-life extension domain and can be used in the compositions and methods disclosed herein.

[0167] Half-life extended structural domain

[0168] In some embodiments, the fusion protein disclosed herein includes a half-life extension domain. In some embodiments, the half-life extension domain increases the cycling half-life of the IGBBPL1 peptide or a fragment thereof.

[0169] Half-life extension domains are well known in the art; see, for example, Strohl, BioDrugs. 2015; 29(4):215-239 and Zaman. people, Journal of Controlled Release 301 (2019) 176-189, for examples of half-life extension domains that can be used in the fusion proteins described herein, are each incorporated herein in their entirety. Half-life extension domains are also described in WO 2016 / 025647, which is incorporated herein by reference below.

[0170] Half-life extension domains that can be used in the fusion proteins described herein include any protein domain that can fuse with the IGBBPL1 domain to extend its half-life. Examples of half-life extension domains include Fc domains, albumin domains, transferrin domains, C-terminal peptides (e.g., the C-terminal peptide of the β chain of human chorionic gonadotropin), elastin-like peptides, inaccurate repeating peptide sequences, proline-alanine-serine polymers, repeating sequences of glycine-rich polypeptides, and gelatin-like proteins.

[0171] Fc structural domain

[0172] In some embodiments, the extended half-life portion is the Fc domain of an immunoglobulin. Various Fc domain gene sequences (e.g., mouse and human constant region gene sequences) are available in the form of publicly available deposits. Constant region domains containing Fc domain sequences can be selected that lack specific effector functions and / or have specific modifications to reduce immunogenicity. Numerous sequences of antibodies and antibody-encoding genes have been disclosed, and suitable Fc domain sequences (e.g., hinge, CH2, and / or CH3 sequences or portions thereof) can be derived from these sequences using techniques recognized in the art. The genetic material obtained using any of the foregoing methods can then be altered or synthesized to obtain a polypeptide suitable for the methods disclosed herein. It should also be understood that the scope of the invention covers alleles, variants, and mutations of constant region DNA sequences.

[0173] The extended half-life domain can be the Fc domain of any type of immunoglobulin, including IgA, IgD, IgE, IgG, and IgM. The extended half-life portion can be the Fc domain of any immunoglobulin subtype, including IgG1, IgG2, IgG3, or IgG4. In some embodiments, the extended half-life domain is the Fc domain of human IgG1.

[0174] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 4. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 4 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0175] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 5. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 5. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 5 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0176] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 6. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 6 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0177] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 7. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 7. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 7 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0178] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 8. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 8. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 8 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0179] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 9. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 9. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 9 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0180] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 137. In some embodiments, the Fc domain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 137. In some embodiments, the Fc domain comprises the sequence shown in SEQ ID NO: 137 with 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0181] In some embodiments, the Fc domain used in the fusion proteins disclosed herein is altered or modified, for example, through amino acid mutations (e.g., addition, deletion, or substitution). As used herein, the term "Fc domain variant" refers to an Fc domain that has at least one amino acid modification (such as amino acid substitution) compared to the wild-type Fc domain from which the Fc domain is derived. For example, where the Fc domain is derived from a human IgG1 antibody, the variant contains at least one amino acid mutation (e.g., substitution) compared to the wild-type amino acid at the corresponding position in the human IgG1 Fc region.

[0182] In some embodiments, the Fc variant contains substitutions at amino acid positions located in the hinge domain or a portion thereof. In some embodiments, the Fc variant contains substitutions at amino acid positions located in the CH2 domain or a portion thereof. In some embodiments, the Fc variant contains substitutions at amino acid positions located in the CH3 domain or a portion thereof. In some embodiments, the Fc variant contains substitutions at amino acid positions located in the CH4 domain or a portion thereof.

[0183] In some embodiments, the fusion protein comprises an Fc variant containing more than one amino acid substitution. The fusion protein may contain, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. Preferably, the amino acid substitutions are spatially separated by at least one amino acid position or more, for example, by at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid positions or more. More preferably, the engineered amino acids are spatially separated by at least 5, 10, 15, 20, or 25 amino acid positions or more.

[0184] In some embodiments, the Fc domain includes variations in the region between amino acids 234-238, including the sequence LLGGP at the start of the CH2 domain. In some embodiments, Fc variants alter Fc-mediated effector functions, particularly ADCC, and / or reduce binding affinity to the Fc receptor. In some embodiments, sequence changes closer to the CH2-CH3 linker at positions such as K322 or P331 can eliminate complement-mediated cytotoxicity and / or alter FcR binding affinity. In some embodiments, the Fc domain incorporates variations at residues P238 and P331, for example, replacing wild-type proline at these positions with serine. In some embodiments, changes in the CCC, SCC, SSC, SCS, or SSS hinge regions encoding one or more of the three hinge cysteine ​​residues can also affect FcR binding and molecular homogeneity, for example, by eliminating unpaired cysteine ​​residues that may destabilize the folded protein.

[0185] Consider other amino acid mutations in the Fc domain to reduce binding to Fc γ receptors and Fc γ receptor subtypes. For example, the locations of region Fc are 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 312, 315, 322, 324, 327, 329, 330, 331, 333, 334, 335, 337, 338, 340, 356, 360. Mutations at positions 373, 376, 378, 379, 382, ​​388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438, or 439 can alter binding, as described in U.S. Patent No. 6,737,056, issued May 18, 2004, which is incorporated herein by reference in its entirety. That patent reports that changing Pro331 in IgG3 to Ser results in a one-sixth reduction in affinity compared to unmutated IgG3, thereby directing Pro331 to participate in FcγRI binding. Furthermore, the amino acid modifications at positions 234, 235, 236 and 237, 297, 318, 320 and 322 are disclosed in US5,624,821, which was authorized on April 29, 1997 and is incorporated herein by reference in its entirety, as potentially altering receptor binding affinity.

[0186] In some embodiments, the Fc domain contains an amino acid substitution at N297. In some embodiments, the amino acid substitution is N297D. In some embodiments, the amino acid substitution is N297Q. In some embodiments, the Fc domain with the N297D mutation contains the amino acid sequence shown in SEQ ID NO: 92. In some embodiments, the Fc domain with the N297Q mutation contains the amino acid sequence shown in SEQ ID NO: 93.

[0187] Other mutations considered for use include, for example, those described in U.S. Patent Application Publication No. 2006 / 0235208, disclosed October 19, 2006 and incorporated herein by reference in its entirety. This disclosure describes Fc variants exhibiting reduced binding to the Fcγ receptor, reduced antibody-dependent cell-mediated cytotoxicity, or reduced complement-dependent cytotoxicity, comprising at least one amino acid modification in the Fc region, including 232G, 234G, 234H, 235D, 235G, 235H, 2361, 236N, 236P, 236R, 237K, 237L, 237N, 237P, 238K, 239R, 265G, 267R, 269R, 270H, 297S, 299A, 2991, 299V, 325A, 325L, 32 7R, 328R, 329K, 3301, 330L, 330N, 330P, 330R and 331L (numbered according to the EU index), and double mutants 236R / 237K, 236R / 325L, 236R / 328R, 237K / 325L, 237K / 328R, 325L / 328R, 235G / 236R, 267R / 269R, 234G / 235G, 236R / 237K / 325L, 236R / 325L / 328R, 235G / 236R / 237K and 237K / 325L / 328R. Other mutations considered for use as described in this disclosure include 227G, 234D, 234E, 234G, 2341, 234Y, 235D, 2351, 235S, 236S, 239D, 246H, 255Y, 258H, 260H, 2641, 267D, 267E, 268D, 268E, 272H, 2721, 272R, 281D, 282G, 283H, 284E, 293R, 295E, 304T, 324G, 3241, 327D, 327A, 328A, 328D, 328E, 328F, 3281, and 328M. 328N, 328Q, 328T, 328V, 328Y, 3301, 330L, 330Y, 332D, 332E,335D Insert G between positions 235 and 236, Insert A between positions 235 and 236, Insert S between positions 235 and 236, Insert T between positions 235 and 236, Insert N between positions 235 and 236, Insert D between positions 235 and 236, Insert V between positions 235 and 236, Insert L between positions 235 and 236, Insert G between positions 235 and 236, Insert A between positions 235 and 236, Insert S between positions 235 and 236, Insert T between positions 235 and 236... Insert N, D between positions 235 and 236, V between positions 235 and 236, L between positions 235 and 236, G between positions 297 and 298, A between positions 297 and 298, S between positions 297 and 298, D between positions 297 and 298, G between positions 326 and 327, A between positions 326 and 327, T between positions 326 and 327, D between positions 326 and 327, and E between positions 326 and 327 (numbered according to the EU index). Additionally, the mutations described in U.S. Patent Application Publication No. 2006 / 0235208 include 227G / 332E, 234D / 332E, 234E / 332E, 234Y / 332E, 2341 332E, 234G / 332E, 235I7332E, 235S / 332E, 235D / 332E, 235E / 332E, 236S / 332E, 236A / 332E, 236S / 332D, 236A / 332D, 239D / 268E, 246H / 332E, 255Y / 332E, 258H / 332E, 260H / 332E, and 2641 332E. 267E / 332E, 267D / 332E, 268D / 332D, 268E / 332D,268E / 332E, 268D / 332E, 268E / 330Y, 268D / 330Y, 272R / 332E, 272H / 332E, 283H / 332E,284E / 332E, 293R / 332E, 295E / 332E, 304T / 332E, 3241 332E, 324G / 332E, 324I7332D,324G / 332D, 327D / 332E, 328A / 332E, 328T / 332E, 328V / 332E, 3281 332E, 328F / 332E, 328Y / 332E, 328M / 332E, 328D / 332E, 328E / 332E, 328N / 332E,328Q / 332E, 328A / 332D, 328T / 332D, 328V / 332D, 3281 332D, 328F / 332D, 328Y / 332D, 328M / 332D, 328D / 332D, 328E / 332D, 328N / 332D, 328Q / 332D, 330L / 332E, 330Y / 332E, 3301 332E, 332D / 330Y, 335D / 332E, 239D / 332E, 239D / 332E / 330Y, 239D / 332E / 330L, 239D / 332E / 330I 239D / 332E / 268E, 239D / 332E / 268D, 239D / 332E / 327D, 239D / 332E / 284E, 239D / 268E / 330Y, 239D / 332E / 268E / 330Y, 239D / 332E / 327A, 239D / 332E / 268E / 327A, 239D / 332E / 330Y / 327A, 332E / 330Y / 268E / 327A, 239D / 332E / 268E / 330Y / 327AConsider using inserts G>297-298 / 332E, insert A>297-298 / 332E, insert S>297-298 / 332E, insert D>297-298 / 332E, insert G>326-327 / 332E, insert A>326- 327 / 332E, Insert T>326-327 / 332E, Insert D>326-327 / 332E, Insert E>326-327 / 332E, Insert G>235-236 / 332E, Insert A>235-236 / 332E, Insert S>235-236 / 332E, Insert T>235-236 / 332E, Insert N>235-236 / 332E, Insert D>235-236 / 332E, Insert V>235-236 / 332E, Insert L>235-236 / 332E, Insert G>235-236 / 332D, Insert A>235-236 / 332D, Insert S>235- 236 / 332D, insert T>235-236 / 332D, insert N>235-236 / 332D, insert D>235-236 / 332D, insert V>235-236 / 332D, and insert L>235-236 / 332D (numbered according to the EU index). The mutants L234A / L235A are described in, for example, U.S. Patent Application Publication No. 2003 / 0108548, disclosed on, for example, June 12, 2003, and incorporated herein by reference in its entirety. In some embodiments, the described modifications are included, alone or in combination. In some embodiments, the mutation is D265A in human IgG1.

[0188] In some embodiments, the Fc domain includes mutations L234A and L235A, with numbering relative to the Kabat numbering scheme of IgG. In some embodiments, the Fc domain includes mutations M252Y, S254T, and T256E, with numbering relative to the Kabat numbering scheme of IgG.

[0189] In some implementations, the fusion protein contains amino acid substitutions for the Fc domain, which alter the antigen-independent effector function of the peptide, particularly the cyclic half-life of the peptide.

[0190] In some embodiments, the fusion proteins disclosed herein comprise Fc variants containing amino acid substitutions that alter the antigen-dependent effector function of the peptide (particularly ADCC or complement activation) compared to the wild-type Fc region. Such fusion proteins exhibit reduced binding to FcRγ compared to the wild-type peptide, and thus mediate reduced effector function. Fc variants with reduced FcRγ binding affinity are expected to reduce effector function, and such molecules could also be used, for example, to treat conditions where undesirable target cell destruction is desired, such as where normal cells express the target molecule, or where prolonged administration of the peptide may lead to undesirable immune system activation.

[0191] In some embodiments, the fusion protein exhibits altered binding to activated FcyR (e.g., Fcyl, Fcylla, or FcyRIIIa). In some embodiments, the fusion protein exhibits altered binding affinity to inhibitory FcyR (e.g., FcyRIIb). Exemplary amino acid substitutions that alter FcR or complement-binding activity are disclosed in International PCT Publication WO05 / 063815, which is incorporated herein by reference.

[0192] The fusion proteins disclosed herein may also include amino acid substitutions that alter the glycosylation of extended PK IL-2. For example, the Fc domain of the fusion protein may include a mutated Fc domain that results in reduced glycosylation (e.g., N-linked or O-linked glycosylation), or may include a modified glycoform (e.g., a low-fucose or fucose-free glycan) of the wild-type Fc domain. In some embodiments, the fusion protein has amino acid substitutions near or within a glycosylation motif, such as an N-linked glycosylation motif containing the amino acid sequence NXT or NXS. Exemplary amino acid substitutions that reduce or alter glycosylation are disclosed in WO05 / 018572 and US2007 / 0111281, the contents of which are incorporated herein by reference. In some embodiments, the fusion proteins disclosed herein include at least one Fc domain having an engineered cysteine ​​residue or the like, located on a solvent-exposed surface. In some embodiments, the fusion protein disclosed herein comprises an Fc domain containing at least one engineered free cysteine ​​residue or its analogue, which is substantially free of disulfide bonds to a second cysteine ​​residue. Any of the aforementioned engineered cysteine ​​residues or their analogues may then be conjugated to a functional domain (e.g., to a thiol-reactive heterobifunctional linker) using techniques well-known in the art.

[0193] In some embodiments, the fusion protein disclosed herein may comprise a genetically fused Fc domain having two or more constituent Fc domains independently selected from the Fc domains described herein. In some embodiments, the Fc domains are identical. In some embodiments, at least two of the Fc domains are different. For example, the Fc domains of the fusion protein disclosed herein contain the same number of amino acid residues, or their lengths may differ by one or more amino acid residues (e.g., by about 5 amino acid residues (e.g., 1, 2, 3, 4, or 5 amino acid residues), about 10 residues, about 15 residues, about 20 residues, about 30 residues, about 40 residues, or about 50 residues). In some embodiments, the Fc domains of the fusion protein disclosed herein may have sequences that differ at one or more amino acid positions. For example, at least two of the Fc domains may differ at about 5 amino acid positions (e.g., 1, 2, 3, 4 or 5 amino acid positions), about 10 positions, about 15 positions, about 20 positions, about 30 positions, about 40 positions or about 50 positions.

[0194] Polyethylene glycolation

[0195] In some embodiments, the half-life extension domain is a polyethylene glycol (PEG) domain. PEGylation is well known in the art to impart an increased cycling half-life to a protein. Methods of PEGylation are well known and disclosed, for example, in US7,610,156 and US7,847,062, all of which are incorporated herein by reference.

[0196] PEG is a well-known water-soluble polymer, commercially available or prepared by ring-opening polymerization of ethylene glycol according to methods well known in the art (Sandler and Karo, Polymer Synthesis, Academic Press, New York, Vol. 3, pp. 138-161). The term "PEG" is broadly used to encompass any polyethylene glycol molecule, regardless of size or modification of the PEG end, and can be represented by the following formula: X— 0(CH2CH2O) n-1CH2CH2OH, where n is 20 to 2300, and X is H or a terminal modification, such as Ci-4 alkyl. In some embodiments, the PEG suitable for the methods disclosed herein is capped at one end with a hydroxyl or methoxy group, i.e., X is H or CH3 (“methoxyPEG”). PEG may contain other chemical groups necessary for the binding reaction; they are produced by the chemical synthesis of the molecule; or they are spacers for optimal distances between the parts of the molecule. Furthermore, such PEG may consist of one or more PEG side chains linked together. PEG having more than one PEG chain is referred to as multi-arm or branched PEG. Branched PEG can be prepared, for example, by adding polyethylene oxide to various polyols, including glycerol, pentaerythritol, and sorbitol. For example, a tetra-arm branched PEG can be prepared from pentaerythritol and ethylene oxide. Branched PEG is described, for example, in EP-A 0 473 084 and US 5,932,462, both of which are incorporated herein by reference. One form of PEG comprises two PEG side chains (PEG2) linked via a primary amino group of lysine (Monfardini et al., BioconjugateChem 1995;6:62-9).

[0197] In some implementations, PEGylated IGBBPL1 is produced via site-directed PEGylation, particularly by conjugation of PEG to a cysteine ​​moiety at the N-terminus or C-terminus. The PEG moiety can also be linked by other chemical methods, including conjugation with an amine. PEG conjugation to a peptide or protein typically involves the activation of PEG and the direct coupling of the activated PEG-intermediate to the target protein / peptide or to a linker, which is subsequently activated and coupled to the target protein / peptide (see Abuchowski et al., JBC 1977;252:3571 and JBC 1977;252:3582, and Harris et al., Poly(ethylene glycol) Chemistry: Biotechnical and Biomedical Applications; (edited by JM Harris) Plenum Press: New York, 1992; Chapters 21 and 22). PEGs of various molecular weights, from about 1,000 Daltons (Da) to 100,000 Da (n is 20 to 2300), can be selected for conjugation with IGBBPL1. The number of repeating units “n” in the PEG approximates the molecular weight described in Daltons. Preferably, the combined molecular weight of the PEGs on the activated linker is suitable for pharmaceutical use. Thus, in one embodiment, the molecular weight of the PEG molecules does not exceed 100,000 Da. For example, if three PEG molecules are conjugated to the linker, each having the same molecular weight of 12,000 Da (each n is about 270), the total molecular weight of the PEGs on the linker is about 36,000 Da (total n is about 820). The molecular weight of the PEGs conjugated to the linker can also be different; for example, of the three molecules on the linker, two PEG molecules can each be 5,000 Da (each n is about 110), and one PEG molecule can be 12,000 Da (n is about 270).

[0198] Those skilled in the art can select an appropriate molecular weight of PEG based, for example, on how PEGylated IGBFPL1 will be used in therapy, the required dose, cycle time, resistance to proteolysis, immunogenicity, and other considerations. For a discussion of PEG and its use in enhancing the properties of proteins, see NV Katre, Advanced Drug Delivery Reviews 1993;10:91-114.

[0199] In some implementations, the PEG molecule can be activated to react with an amino group on IL-2, such as with lysine (Bencham CO et al., Anal. Biochem., 131, 25 (1983); Veronese, FM et al., Appl. Biochem., 11, 141 (1985); Zalipsky, S. et al., Polymeric Drugs and Drug Delivery Systems, adrs 9-110 ACS Symposium Series 469 (1999); Zalipsky, S. et al., Europ. Polym. J., 19, 1177-1183 (1983); Delgado, C. et al., Biotechnology and Applied Biochemistry, 12, 119-128 (1990)).

[0200] In some embodiments, PEG carbonates are used to form PEG-IL-2 conjugates. N,N'-disuccinimidyl carbonate (DSC) can be used in the reaction with PEG to form an active mixed PEG-succinimidyl carbonate, which can then be reacted with the nucleophilic group of the linker or the amino group of IL-2 (see U.S. Patent Nos. 5,281,698 and 5,932,462). In similar types of reactions, 1,1'-(dibenzotriazolyl) carbonate and bis-(2-pyridyl) carbonate can be reacted with PEG to form PEG-benzotriazolyl and PEG-pyridyl mixed carbonates, respectively (U.S. Patent No. 5,382,657). PEGylation of IGFBPL1 can be carried out according to prior art methods, such as by reacting IGFBPL1 with electrophilic active PEG (Shearwater Corp., USA, www.shearwatercorp.com). Preferred PEG reagents suitable for use in the methods disclosed herein are, for example, N-hydroxysuccinimide propionate (PEG-SPA), butyrate (PEG-SBA), PEG-succinimide propionate, or branched N-hydroxysuccinimide (such as mPEG2-NHS) (Monfardini, C et al., Bioconjugate Chem. 6 (1995) 62-69).

[0201] In some implementations, PEGylated IGBBPL1 comprises one or more PEG molecules covalently linked to the connector.

[0202] Other half-life extended domains

[0203] In some embodiments, the half-life extension domain is serum albumin or a fragment thereof. Methods of fusing serum albumin with proteins are disclosed in, for example, US2010 / 0144599, US2007 / 0048282, and US2011 / 0020345, which are incorporated herein by reference in their entirety. In some embodiments, the half-life extension domain is human serum albumin (HSA) or a variant or fragment thereof, such as those disclosed in US 5,876,969, WO 2011 / 124718, WO 2013 / 075066, and WO 2011 / 0514789.

[0204] In some embodiments, serum albumin comprises the amino acid sequence of SEQ ID NO: 181. In some embodiments, serum albumin comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the sequence shown in SEQ ID NO: 181. In some embodiments, serum albumin comprises the sequence shown in SEQ ID NO: 181 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0205] In some embodiments, the half-life extension domain is a serum albumin-binding protein, such as those described in US2005 / 0287153, US2007 / 0003549, US2007 / 0178082, US2007 / 0269422, US2010 / 0113339, WO2009 / 083804 and WO2009 / 133208, which are incorporated herein by reference in their entirety.

[0206] In some implementations, the half-life extension domain is a transferrin, as disclosed in US 7,176,278 and US 8,158,579, which are incorporated herein by reference in their entirety.

[0207] In some implementations, the half-life extension domain is a serum immunoglobulin-binding protein, such as those disclosed in US2007 / 0178082, which is incorporated herein by reference in its entirety.

[0208] In some embodiments, the half-life extension domain is a fibronectin (Fn)-based scaffold domain protein that binds to serum albumin, such as those disclosed in US2012 / 0094909, which is incorporated herein by reference in its entirety. Methods for preparing fibronectin-based scaffold domain proteins are also disclosed in US2012 / 0094909. A non-limiting example of an Fn3-based extended PK group is Fn3(HSA), i.e., the Fn3 protein that binds to human serum albumin.

[0209] connector

[0210] In some implementations, the IGBBPL1 peptide or a fragment thereof is operatively linked to a half-life extension domain via a linker.

[0211] Linkers that can be used to connect domains of fusion proteins are well known in the art; see, for example, Chen et al., Adv Drug Deliv Rev. 2013 Oct;65(10):1357-69.

[0212] In some embodiments, the IGBBPL1 peptide or a fragment thereof and the half-life extension domain are linked via a synthetic linker. As used herein, the term "synthetic" in relation to a peptide linker includes a peptide (or polypeptide) comprising an amino acid sequence (which may or may not be naturally occurring) linked in a linear amino acid sequence to a sequence in nature that is not naturally linked (which may or may not be naturally occurring) (e.g., an ABP sequence). For example, a peptide linker may comprise a non-naturally occurring polypeptide that is a modified form of a naturally occurring polypeptide (e.g., containing mutations such as addition, substitution, or deletion) or that comprises a first amino acid sequence (which may or may not be naturally occurring).

[0213] In some embodiments, the IGBBPL1 peptide or a fragment thereof and the extended half-life domain are linked via a cleavable linker. Examples of cleavable linkers include cyclic peptide linkers, disulfide linkers, and linkers containing sequences susceptible to protease cleavage. In some embodiments, the IGBBPL1 peptide or a fragment thereof and the extended half-life domain are linked via a linker containing a tobacco etch virus (TEV) cleavage site. An illustrative sequence of the TEV cleavable linker is GGGSENLYFQS (SEQ ID NO: 12). In some embodiments, the TEV cleavable linker comprises the amino acid sequence of SEQ ID NO: 182.

[0214] In some embodiments, the IGBBPL1 peptide or a fragment thereof and the half-life extension domain are linked by a flexible linker. The flexible linker typically contains a small nonpolar amino acid (such as glycine) or a polar amino acid (such as serine and / or threonine). An example of a flexible linker is the GS linker, such as the G4S linker. The G4S linker contains the formula (Gly4Ser). nThe amino acid sequence, where n is a positive integer (e.g., 1, 2, 3, 4, or 5). In some embodiments, the gly / ser adapter is (Gly4Ser)1. In some embodiments, the gly / ser adapter is (Gly4Ser)2. In some embodiments, the gly / ser adapter is (Gly4Ser)3. In some embodiments, the gly / ser adapter is (Gly4Ser)4. In some embodiments, the gly / ser adapter is (Gly4Ser)5. In some embodiments, two or more gly-ser adapters are tandemly incorporated into the peptide adapter.

[0215] In some embodiments, the IGBBPL1 peptide or a fragment thereof and the half-life extension domain are linked via a G4S linker. In some embodiments, the IGBBPL1 peptide or a fragment thereof and the half-life extension domain are linked via a GS linker. Illustrative sequences of the GS linker include GGGGSGGGGSGGGGS (SEQ ID NO:13) and GGGSGGSGGSS (SEQ ID NO:14). In some embodiments, the GS linker comprises the amino acid sequence of SEQ ID NO: 159.

[0216] Other linkers suitable for use in the fusion proteins described herein are known in the art, such as the serine-rich linker disclosed in US 5,525,491; the helical-forming peptide linker (e.g., A(EAAAK)nA (n=2-5) (SEQ ID NO:23)) disclosed in Arai et al., Protein Eng 2001;14:529-32; and the stable linker, namely the dipeptide linker LE, disclosed in Chen et al., Mol Pharm 2011;8:457-65; the thrombin-sensitive disulfide cyclic peptide linker; and the α-helical-forming linker LEA(EAAAK)4ALEA(EAAAK)4ALE (SEQ ID NO:24).

[0217] In some embodiments, the adapter is the XTEN-13k adapter. In some embodiments, the XTEN-13k adapter contains the amino acid sequence of SEQ ID NO: 180.

[0218] Fusion protein structure and exemplary fusion proteins

[0219] The IGFBPL1 peptide or a fragment thereof and the half-life extension domain may be arranged in any order in the fusion protein provided herein. The components for binding DCC, SORT1, or both DCC and SORT1, and the half-life extension domain may be arranged in any order in the fusion protein provided herein. In some embodiments, the IGFBPL1 peptide or a fragment thereof is located at the N-terminus of the half-life extension domain. In some embodiments, the IGFBPL1 peptide or a fragment thereof is located at the C-terminus of the half-life extension domain.

[0220] In some embodiments, the two fusion proteins described herein dimerize. It will be apparent to those skilled in the art that the sequence of the fusion protein domains can be manipulated to stabilize dimerization. For example, in some embodiments, a pestle-and-mortar design can be used to ensure the dimerization of the fusion protein. SEQ ID No:5 and 8 are exemplary "pestle" Fc sequences, and SEQ ID No:6 and 9 are exemplary "mortar" Fc sequences. Methods for implementing pestle-and-mortar modifications are known in the art; see, for example, Liu... wait people Front. Immunol., January 26, 2017, Chapter 2 Vaccines and Molecular Therapeutics, Volume 8 - 2017.

[0221] The fusion protein dimers described herein can be monovalent or multivalent. Each IGBBPL1 polypeptide or fragment thereof in a multivalent fusion protein dimer has more than one extended half-life portion.

[0222] In some embodiments, the fusion protein dimer is a monovalent fusion protein dimer. In some embodiments, the fusion protein dimer is a monovalent N-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a monovalent C-terminal fusion protein dimer.

[0223] In some embodiments, the fusion protein dimer is a divalent fusion protein dimer. In some embodiments, the fusion protein dimer is a divalent N-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a divalent C-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a divalent N-terminal and C-terminal fusion protein dimer.

[0224] In some embodiments, the fusion protein is a trivalent fusion protein dimer. In some embodiments, the fusion protein dimer is a trivalent N-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a trivalent C-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a trivalent N-terminal and C-terminal fusion protein dimer.

[0225] In some embodiments, the fusion protein is a tetravalent fusion protein dimer. In some embodiments, the fusion protein dimer is a tetravalent N-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a tetravalent C-terminal fusion protein dimer. In some embodiments, the fusion protein dimer is a tetravalent N-terminal and C-terminal fusion protein dimer.

[0226] If the fusion protein dimer is a divalent fusion protein, the half-life extension domain can be linked in either a trans or cis orientation. In some embodiments, the fusion protein is a divalent N-terminal and C-terminal trans fusion protein. In some embodiments, the fusion protein is a divalent N-terminal and C-terminal cis fusion protein.

[0227] In some embodiments, the fusion protein is a monovalent N-terminal Fc fusion protein. In some embodiments, the fusion protein is a monovalent C-terminal Fc fusion protein. In some embodiments, the fusion protein is a bivalent N-terminal Fc fusion protein. In some embodiments, the fusion protein is a bivalent C-terminal Fc fusion protein. In some embodiments, the fusion protein is a tetravalent N-terminal and C-terminal trans Fc fusion protein. In some embodiments, the fusion protein is a bivalent N-terminal and C-terminal cis Fc fusion protein.

[0228] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 15. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 15. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 15 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0229] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 16. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 16. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 16 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0230] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 17. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 17. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 17 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0231] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 18. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 18. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 18 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0232] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 19. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 19. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 19 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0233] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 20. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 20. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 20 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0234] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 21. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 21. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 21 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0235] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 22. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 22. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 22 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0236] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 51. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 51. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 51 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0237] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 52. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 52 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0238] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 53. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 53. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 53 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0239] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 54. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 54. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 54 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0240] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 55. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 55 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0241] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 56. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 56 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0242] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 57. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 57. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 57 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0243] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 58. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 58 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0244] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 59. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 59 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0245] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 60. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 60 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0246] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 61. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 61. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 61 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0247] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 62. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 62. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 62 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0248] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 63. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 63. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 63 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0249] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 64. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 64. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 64 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0250] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 65. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 65. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 65 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0251] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 66. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 66. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 66 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0252] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 67. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 67. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 67 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0253] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 68. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 68. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 68 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0254] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 69. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 69. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 69 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0255] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 70. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 70. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 70 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0256] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 71. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 71. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 71 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0257] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 72. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 72. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 72 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0258] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 73. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 73. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 73 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0259] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 74. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 74. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 74 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0260] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 75. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 75. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 75 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0261] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 76. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 76. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 76 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0262] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 77. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 77. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 77 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0263] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 78. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 78. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 78 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0264] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 79. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 79. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 79 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0265] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 80. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 80. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 80 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0266] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 81. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 81. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 81 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0267] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 82. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 82. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 82 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0268] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 83. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 83. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 83 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0269] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 84. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 84. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 84 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0270] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 85. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 85. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 85 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0271] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 86. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 86. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 86 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0272] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 94. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 94. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 94 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0273] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 95. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 95. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 95 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0274] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 96. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 96. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 96 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0275] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 97. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 97. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 97 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0276] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 98. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 98. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 98 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0277] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 99. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 99. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 99 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0278] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 100. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 100. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 100 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0279] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 101. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 101. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 101 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0280] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 102. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 102. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 102 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0281] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 103. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 103. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 103 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0282] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 104. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 104. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 104 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0283] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 105. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 105. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 105 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0284] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 106. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 106. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 106 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0285] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 107. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 107. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 107 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0286] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 108. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 108. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 108 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0287] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 109. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 109. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 109 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0288] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 110. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 110. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 110 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0289] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 111. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 111. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 111 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0290] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 112. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 112. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 112 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0291] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 113. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 113. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 113 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0292] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 114. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 114. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 114 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0293] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 115. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 115. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 115 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0294] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 116. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 116. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 116 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0295] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 117. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 117. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 117 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0296] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 118. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 118. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 118 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0297] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 119. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 119. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 119 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0298] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 120. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 120. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 120 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0299] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 121. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 121. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 121 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0300] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 122. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 122. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 122 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0301] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 123. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 123. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 123 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0302] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 124. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 124. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 124 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0303] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 125. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 125. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 125 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0304] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 126. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 126. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 126 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0305] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 127. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 127. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 127 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0306] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 128. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 128. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 128 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0307] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 129. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 129. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 129 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0308] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 130. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 130. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 130 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0309] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 131. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 131. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 131 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0310] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 132. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 132. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 132 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0311] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 133. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 133. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 133 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0312] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 134. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 134. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 134 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0313] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 135. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 135. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 135 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0314] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 136. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 136. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 136 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0315] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 138. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 138. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 138 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0316] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 139. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 139. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 139 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0317] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 140. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 140. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 140 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0318] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 142. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 142. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 142 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0319] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 144. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 144. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 144 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0320] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 145. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 145. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 145 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0321] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 146. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 146. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 146 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0322] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 147. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 147. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 147 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0323] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 148. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 148. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 148 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0324] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 149. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 149. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 149 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0325] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 150. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 150. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 150 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0326] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 183. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 183. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 183 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0327] In some embodiments, the fusion protein comprises the amino acid sequence shown in SEQ ID NO: 184. In some embodiments, the fusion protein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 184. In some embodiments, the fusion protein comprises the sequence shown in SEQ ID NO: 184 having 1, 2, 3, 4, or 5 conserved amino acid substitutions.

[0328] In some embodiments, the fusion protein is a dimer comprising two identical fusion proteins described herein. In some embodiments, the fusion protein is a dimer comprising two distinct fusion proteins described herein. In some embodiments, the fusion protein is a dimer comprising the fusion protein described herein and a half-life extension domain lacking the IGBBPL1 polypeptide or a fragment thereof. In some embodiments, the half-life extension domain of one or more fusion proteins contains modifications that induce dimer formation (e.g., a mortar-and-mortar modification of the Fc domain).

[0329] In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 20, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 21. In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 20, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 21.

[0330] In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 22, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 9. In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 22, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 9.

[0331] In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 134, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 21. In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 134, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 21.

[0332] In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 138, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 139. In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 138, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 139.

[0333] In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 140, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 141. In some embodiments, the fusion protein is a dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 140, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 141.

[0334] Methods for preparing peptides

[0335] Methods for preparing peptides are known in the art, as described in WO 2016 / 025647 and provided below. In some embodiments, the peptides and fusion proteins described herein are prepared in transformed host cells using recombinant DNA technology. For this purpose, recombinant DNA molecules encoding peptides are prepared. Methods for preparing such DNA molecules are well known in the art. For example, the sequence encoding the peptide can be excised from DNA using a suitable restriction enzyme. Alternatively, DNA molecules can be synthesized using chemical synthesis techniques such as the aminophosphate method. Moreover, combinations of these techniques can be used.

[0336] Methods for preparing peptides also include vectors capable of expressing the peptide in a suitable host. The vector contains a DNA molecule encoding a peptide operatively linked to an appropriate expression control sequence. Methods for influencing this operative linking before or after the DNA molecule is inserted into the vector are well known. Expression control sequences include promoters, activators, enhancers, operons, ribosomal nuclease domains, initiation signals, termination signals, cap signals, polyadenylation signals, and other signals involved in controlling transcription or translation.

[0337] The resulting vector, containing DNA molecules, is used to transform a suitable host. This transformation can be performed using methods well known in the art.

[0338] Any of the numerous available and well-known host cells is suitable for the methods disclosed herein. The choice of a particular host depends on many factors recognized in the art. These factors include, for example, compatibility with the chosen expression vector, toxicity of the peptide encoded by the DNA molecule, conversion efficiency, ease of peptide recovery, expression characteristics, biosafety, and cost. A balance must be struck among these factors, understanding that not all hosts may be equally effective for expression of a particular DNA sequence. In these general guidelines, useful microbial hosts include bacteria (e.g., *Escherichia coli* sp.), yeasts (e.g., *Saccharomyces* sp.), and other fungi, insects, plants, cultured mammalian (including human) cells, or other hosts known in the art.

[0339] Next, the transformed host cells are cultured and purified. The host cells can be cultured under standard fermentation conditions to express the desired compound. Such fermentation conditions are well known in the art. Finally, the peptides are purified from the culture using methods well known in the art.

[0340] Compounds can also be prepared by synthetic methods. For example, solid-phase synthesis can be used. Suitable techniques are well known in the art, including those described in Merrifield (1973), Chem. Polypeptides, pp. 335-61 (edited by Katsoyannis and Panayotis); Merrifield (1963), J. Am. Chem. Soc. 85: 2149; Davis et al. (1985), Biochem. Intl. 10: 394-414; Stewart and Young (1969), Solid Phase Peptide Synthesis; U.S. Patent No. 3,941,763; Finn et al. (1976), The Proteins (3rd Edition) 2: 105-253; and Erickson et al. (1976), The Proteins (3rd Edition) 2: 257-527. Solid-phase synthesis is the preferred technique for preparing single peptides because it is the most cost-effective method for preparing small peptides.

[0341] Compounds containing derivatized peptides or non-peptide groups can be synthesized using well-known organic chemistry techniques.

[0342] Other methods of molecular expression / synthesis are generally known to those skilled in the art.

[0343] The aforementioned nucleic acid molecules can be contained within a vector that directs their expression in cells, for example, cells already transduced with the vector. Therefore, in some embodiments, in addition to polypeptide mutants, expression vectors containing nucleic acid molecules encoding mutants and cells transfected with these vectors are also available.

[0344] Suitable vectors include T7-based vectors for bacteria (see, for example, Rosenberg et al., Gene 56: 125, 1987), pMSXND expression vectors for mammalian cells (Lee and Nathans, J. Biol. Chem. 263:3521, 1988), and baculovirus-derived vectors for insect cells (e.g., pBacPAKS expression vector from Clontech, Palo Alto, Calif.). The nucleic acid insert encoding the polypeptide of interest in such vectors can be operatively linked to a promoter selected based on, for example, the cell type for which expression is sought. For example, the T7 promoter can be used in bacteria, the polyhedrosis protein promoter in insect cells, and cytomegalovirus or metallothionein promoters in mammalian cells. Furthermore, in the case of higher eukaryotes, tissue-specific and cell-type-specific promoters are widely available. These promoters are named for their ability to direct the expression of nucleic acid molecules in a given tissue or cell type in vivo. Many promoters and other regulatory elements that can be used to direct nucleic acid expression are well known to those skilled in the art.

[0345] In addition to sequences that facilitate transcription of the inserted nucleic acid molecules, vectors can also contain other genes encoding origin of replication and selectivity markers. For example, neomycin resistance (neomycin resistance...) r The gene confers G418 resistance to cells that express it, thereby allowing phenotypic selection of transfected cells. Those skilled in the art can readily determine whether a given regulatory element or selection marker is suitable for a particular experimental setting.

[0346] Suitable viral vectors include, for example, retroviruses, adenoviruses and adeno-associated vectors, herpesviruses, simian virus 40 (SV40) and bovine papillomavirus vectors (see, for example, Gluzman (ed.), Eukaryotic Viral Vectors, CSH Laboratory Press, Cold Spring Harbor, NY).

[0347] Prokaryotic or eukaryotic cells containing and expressing nucleic acid molecules encoding mutant polypeptides are also suitable for use. The cells are transfected cells, i.e., cells in which nucleic acid molecules (e.g., nucleic acid molecules encoding mutant polypeptides) have been introduced via recombinant DNA technology. Progeny of such cells are also considered suitable for the methods disclosed herein.

[0348] The precise composition of the expression system is not critical. For example, peptide mutants can be generated in prokaryotic hosts (such as the bacteria *Escherichia coli*) or eukaryotic hosts (such as insect cells (e.g., Sf21 cells) or mammalian cells (e.g., COS cells, NIH 3T3 cells, or HeLa cells)). These cells are available from many sources, including the American Type Culture Collection (Manassas, Va.). When selecting an expression system, only the compatibility of the components with each other is important. This determination can be made by a technician or a person with general technical skills. Furthermore, if guidance is needed in selecting an expression system, a technician can consult Ausubel et al. (Current Protocols in Molecular Biology, John Wiley and Sons, New York, NY, 1993) and Pouwels et al. (Cloning Vectors: A Laboratory Manual, 1985 Supplement 1987).

[0349] The expressed peptides can be purified from the expression system using conventional biochemical procedures and can be used as, for example, therapeutic agents, as described herein.

[0350] Pharmaceutical Composition

[0351] This disclosure provides pharmaceutical compositions of the IGBBPL1 peptides and fusion proteins described herein with pharmaceutically acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants; pharmaceutical compositions comprising cancer vaccines with pharmaceutically acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants; pharmaceutical compositions comprising therapeutic antibodies with pharmaceutically acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants; or pharmaceutical compositions comprising immune checkpoint inhibitors with pharmaceutically acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants.

[0352] In some embodiments, the pharmaceutical composition comprises the IGBBPL1 peptide or fusion protein described herein, and pharmaceutically acceptable diluents, carriers, solubilizers, emulsifiers, preservatives, and / or adjuvants. In some embodiments, the acceptable formulation material is preferably non-toxic to the recipient at the dose and concentration used. In some embodiments, the formulation material is intended for subcutaneous (SC) and / or intravenous (IV) administration. In some embodiments, the pharmaceutical composition may contain formulation materials for altering, maintaining, or preserving, for example, the composition's pH, osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeation. In some embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobial agents; antioxidants (such as ascorbic acid, sodium sulfite, or sodium bisulfite); buffers (such as borates, bicarbonates, Tris-HCl, citrates, phosphates, or other organic acids); fillers (such as mannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose, or dextrin); proteins (such as serum albumin, gelatin, or immunoglobulins); colorants, flavorings, and diluents; emulsifiers; hydrophilic polymers (such as polyvinylpyrrolidone); and low molecular weight polymers. Peptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide); solvents (such as glycerol, propylene glycol, or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, dehydrated sorbitol esters, polysorbate esters (such as polysorbate 20, polysorbate 80), triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancers (such as sucrose or sorbitol); tension enhancers (such as alkali metal halides, preferably sodium chloride or potassium chloride, mannitol, sorbitol); delivery media; diluents; excipients and / or pharmaceutical adjuvants. (Remington's Pharmaceutical Sciences, 18th edition, edited by AR Gennaro, Mack Publishing Company (1995)). In some embodiments, the formulation contains PBS; 20 mM NaOAC, pH 5.2, 50 mM NaCl; and / or 10 mM NaOAC, pH 5.2, 9% sucrose.In some embodiments, the optimal pharmaceutical composition will be determined by those skilled in the art based on, for example, the intended route of administration, delivery form, and required dosage. See, for example, Remington's Pharmaceutical Sciences, ibid. In some embodiments, such compositions may affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the IGBBPL1 peptide or fusion protein.

[0353] In some embodiments, the primary medium or carrier in the pharmaceutical composition may be aqueous or non-aqueous in nature. For example, in some embodiments, a suitable medium or carrier may be water for injection, physiological saline solution, or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. In some embodiments, the saline includes isotonic phosphate-buffered saline. In some embodiments, neutral buffered saline or saline mixed with serum albumin are further exemplary mediums. In some embodiments, the pharmaceutical composition comprises a Tris buffer of about pH 7.0-8.5, or an acetate buffer of about pH 4.0-5.5, which may further comprise sorbitol or a suitable alternative thereof. In some embodiments, a composition containing a fusion protein may be prepared for storage in the form of a lyophilized cake or aqueous solution by mixing the selected composition having the desired purity with an optional formulation reagent (Remington's Pharmaceutical Sciences, ibid.). Furthermore, in some embodiments, the composition containing a fusion protein is formulated into a lyophilized product using a suitable excipient (such as sucrose).

[0354] In some embodiments, the pharmaceutical composition may be selected for parenteral delivery. In some embodiments, the composition may be selected for inhalation or for delivery via the digestive tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the capabilities of those skilled in the art.

[0355] In some embodiments, the pharmaceutical composition is formulated for delivery to the eye. In some embodiments, the pharmaceutical composition is formulated for intravitreal administration. In some embodiments, the pharmaceutical composition is formulated for intraocular administration. In some embodiments, the pharmaceutical composition is formulated for subconjunctival administration. In some embodiments, the pharmaceutical composition is formulated for topical application to the eye.

[0356] In some embodiments, the formulation components are present at concentrations acceptable at the application site. In some embodiments, a buffer solution is used to maintain the composition at a physiological pH or slightly lower, typically in the pH range of about 5 to about 8.

[0357] In some embodiments, when considering parenteral administration, the therapeutic composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution containing the fusion protein in a pharmaceutically acceptable medium. In some embodiments, the medium for parenteral injection is sterile distilled water, wherein the fusion protein is formulated into a suitably preserved sterile isotonic solution. In some embodiments, preparation may involve formulating the desired molecule with reagents such as injectable microspheres, biodegradable particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads, or liposomes, which can provide controlled or sustained release of the product, which can then be delivered via accumulation injection. In some embodiments, hyaluronic acid may also be used, and may have an effect that promotes duration of circulation. In some embodiments, an implantable drug delivery device may be used to introduce the desired molecule.

[0358] In some embodiments, the pharmaceutical composition is formulated for inhalation. In some embodiments, the fusion protein is formulated as a dry powder for inhalation. In some embodiments, an inhalation solution containing the fusion protein is formulated together with a propellant for aerosol delivery. In some embodiments, the solution may be nebulized. Lung administration is further described in PCT application PCT / US94 / 001875, which describes the pulmonary delivery of a chemically modified protein.

[0359] In some embodiments, the formulation is intended for oral administration. In some embodiments, the fusion protein administered in this manner may be formulated with or without those carriers typically used to formulate solid dosage forms, such as tablets and capsules. In some embodiments, the capsule may be designed to release the active portion of the formulation at a point in the gastrointestinal tract, where bioavailability is maximized and pre-systemic degradation is minimized. In some embodiments, at least one additional agent is included to facilitate the absorption of the fusion protein. In some embodiments, diluents, flavoring agents, low-melting-point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrants, and binders may also be used.

[0360] In some embodiments, the pharmaceutical composition comprises an effective amount of fusion protein mixed with a non-toxic excipient suitable for manufacturing tablets. In some embodiments, a solution can be prepared in unit dose form by dissolving the tablet in sterile water or another suitable medium. In some embodiments, suitable excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, or sodium bicarbonate, lactose, or calcium phosphate; or binders such as starch, gelatin, or gum arabic; or lubricants such as magnesium stearate, stearic acid, or talc.

[0361] Other pharmaceutical compositions will be apparent to those skilled in the art, including formulations involving the sustained or controlled delivery of fusion proteins in formulations. In some embodiments, techniques for formulating a variety of other sustained or controlled delivery methods, such as liposome carriers, biodegradable microparticles or porous beads, and long-acting injections, are also known to those skilled in the art. See, for example, PCT application PCT / US93 / 00829, which describes the controlled release of porous polymeric microparticles for delivering pharmaceutical compositions. In some embodiments, sustained-release formulations may comprise a semi-permeable polymeric matrix, such as a membrane or microcapsule, in the form of a shaped article. Sustained-release matrices may include polyesters, hydrogels, polylactides (US Patent Nos. 3,773,919 and EP 058,481), copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid esters (Sidman et al., Biopolymers, 22:547-556 (1983)), poly(2-hydroxyethyl-methacrylate) (Langer et al., J. Biomed. Mater. Res., 15: 167-277 (1981) and Langer, Chem. Tech., 12:98-105 (1982)), ethylene vinyl acetate (Langer et al., ibid.), or poly-D(-)-3-hydroxybutyric acid (EP 133,988). In some embodiments, the sustained-release composition may also include liposomes, which may be prepared by any of several methods known in the art. See, for example, Eppstein et al., Proc. Natl. Acad. Sci. USA, 82:3688-3692 (1985); EP 036,676; EP 088,046 and EP 143,949.

[0362] Pharmaceutical compositions intended for internal administration are typically sterile. In some embodiments, this can be accomplished by filtration through a sterile filter membrane. In some embodiments, sterilization using this method may be performed before or after lyophilization and reconstitution when the composition is lyophilized. In some embodiments, compositions intended for parenteral administration may be stored in lyophilized form or in solution. In some embodiments, parenteral compositions are typically placed in containers with sterile access ports, such as intravenous solution bags or vials with stoppers that can be punctured by a hypodermic needle.

[0363] In some embodiments, once the pharmaceutical composition is formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or as a dehydrated or lyophilized powder. In some embodiments, such formulations can be stored in ready-to-use form or in a form reconstituted prior to application (e.g., lyophilized).

[0364] In some embodiments, a kit for generating a single-dose administration unit is provided. In some embodiments, the kit may contain a first container with dried protein and a second container with an aqueous formulation. In some embodiments, kits include pre-filled syringes (e.g., liquid syringes and lyophilized syringes).

[0365] In some embodiments, the effective amount of the pharmaceutical composition containing the fusion protein used in treatment will depend, for example, on the treatment context and purpose. Those skilled in the art will understand that, according to certain embodiments, the appropriate dose level for treatment will therefore vary in part depending on the molecule delivered, the indication for use of the fusion protein, the route of administration, and the patient's size (weight, body surface, or organ size) and / or condition (age and general health status). In some embodiments, clinicians may titrate the dose and modify the route of administration to achieve optimal therapeutic effect.

[0366] In some embodiments, the dosing frequency will take into account the pharmacokinetic parameters of the fusion protein in the formulation used. In some embodiments, the clinician will administer the composition until a dose is reached to achieve the desired effect. In some embodiments, the composition may therefore be administered as a single dose, or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via an implanted device or catheter. Further refinement of the appropriate dose is routinely performed by those skilled in the art and within the scope of their routine duties. In some embodiments, the appropriate dose can be determined by using appropriate dose-response data.

[0367] In some embodiments, the route of administration of the pharmaceutical composition is consistent with known methods, such as oral administration, intravenous, intraperitoneal, intracerebral (parenchymal), intraventricular, intramuscular, subcutaneous, intraocular, intraarterial, portal vein, or intralesional injection; via a continuous release system or via an implantable device. In some embodiments, the composition may be administered continuously by bolus or infusion, or via an implantable device. In some embodiments, the various elements of the combination therapy may be administered via different routes.

[0368] In some embodiments, the composition is administered topically by implanting a membrane, sponge, or other suitable material on which the desired molecule has been absorbed or encapsulated. In some embodiments, when using an implantable device, the device can be implanted into any suitable tissue or organ, and the desired molecule can be delivered by diffusion, timed release bolus, or continuous administration. In some embodiments, it may be necessary to use the pharmaceutical composition fusion protein ex vivo. In this case, cells, tissues, and / or organs removed from the patient are exposed to the pharmaceutical composition containing the fusion protein, and then the cells, tissues, and / or organs are implanted back into the patient.

[0369] Reagent test kit

[0370] In some embodiments, this disclosure provides kits comprising the IGBBPL1 peptide or IGBBPL1 fragment described herein and instructions for use. In some embodiments, this disclosure provides kits comprising a fusion protein and instructions for use. The kit may contain the fusion protein, one or more controls, and various buffers, reagents, enzymes, and other standard components well known in the art in a suitable container. The container may include at least one vial, well, test tube, flask, bottle, syringe, or other container device in which the fusion protein may be placed and, in some cases, appropriately aliquoted. Where additional components are provided, the kit may include additional containers in which such components may be placed. The kit may also include a device for containing the fusion protein, and any other reagent containers tightly limited for commercial sale. Such containers may include injection-molded or blow-molded plastic containers in which the desired vial is held. The container and / or kit may include labels with instructions for use and / or warnings.

[0371] How to use

[0372] In some embodiments, this disclosure provides methods for using the IGBBPL1 peptide described herein. In some embodiments, this disclosure provides methods for using the IGBBPL1 fragment described herein. In some embodiments, this disclosure provides methods for using the fusion protein described herein. In any of the methods described herein, the fusion protein is monovalent, bivalent, trivalent, or tetravalent.

[0373] In some embodiments, this disclosure provides a method for inhibiting SORT1 activity in cells, comprising contacting cells with the IGBBPL1 peptide or IGBBPL1 fragment described herein. In some embodiments, this disclosure provides a method for inhibiting SORT1 activity in cells, comprising contacting cells with the fusion protein described herein. In some embodiments, the cells are microglia. In some embodiments, the microglia are in the brain or eye of a subject. In some embodiments, this disclosure provides a method for increasing the secretion of pro-inflammatory cytokines from cells by contacting cells with the fusion protein described herein. In some embodiments, the cells are microglia. In some embodiments, the cytokines are IL-10, GM-CSF, MIP-1α, or any combination thereof.

[0374] In some embodiments, this disclosure provides a method for inhibiting the secretion of inflammatory cytokines from cells by contacting cells with the IGBBPL1 peptide or IGBBPL1 fragment described herein. In some embodiments, this disclosure provides a method for inhibiting the secretion of inflammatory cytokines from cells by contacting cells with the fusion protein described herein. In some embodiments, the cells are microglia. In some embodiments, the cytokine is IL-6.

[0375] Methods for quantifying proteins are known in the art and include, for example, but not limited to, ELISA and Western blotting.

[0376] In some embodiments, this disclosure provides a method for inhibiting DCC activity in cells, comprising contacting the cells with the IGBBPL1 peptide or IGBBPL1 fragment described herein. In some embodiments, this disclosure provides a method for inhibiting DCC activity in cells, comprising contacting the cells with the fusion protein described herein. In some embodiments, the cells are neurons. In some embodiments, the neurons are in the brain or eye of a subject. In some embodiments, this disclosure provides a method for increasing the number of neurons and / or neurite growth, comprising contacting the neurons with the fusion protein described herein.

[0377] Methods for determining the number of neurons are known to those skilled in the art. For example, cell populations containing neurons can be counted manually. Methods for determining neurite extension are known to those skilled in the art. For example, cell populations containing neurons can be fixed and imaged to visualize neurite extension. Measurements of neurite extension can be determined manually.

[0378] In some embodiments, this disclosure provides a method for inducing phagocytic activity of microglia by contacting them with the IGBBPL1 peptide or IGBBPL1 fragment described herein. In some embodiments, this disclosure provides a method for inducing phagocytic activity of microglia by contacting them with the fusion protein described herein. Methods for determining phagocytic activity are known to those skilled in the art and include using fluorescent latex beads or particles, such as yeast glycans, and subsequently counting the number of cells that take up the beads, wherein an increase in bead uptake indicates an increase in phagocytic activity.

[0379] The IGBBPL1 peptide or fragment described in this article can also be used to preferentially target neurons rather than microglia, and vice versa. The fusion protein described in this article can also be used to preferentially target neurons rather than microglia, and vice versa. Without being bound by theory, it is believed that the two ends of hIGFBPL1 are differentially involved in mediating cell binding, making the two ends of IGBBPL1 important for mediating neuroprotective (C-terminus) and anti-inflammatory (N-terminus) activities. This is thought to be because the N-terminal domain of IGBBPL1 binds to microglia, while the C-terminal domain of IGBBPL1 binds to neurons.

[0380] The IGFFBPL1 peptide or fragment exhibiting "preferential targeting" of neurons rather than microglia shows a stronger binding affinity to neurons than to microglia. In some embodiments, the IGFFBPL1 peptide or fragment that preferentially targets neurons binds to neurons with an affinity at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 25 times that of microglia.

[0381] Fusion proteins or fusion protein dimers that exhibit "preferential targeting" of neurons rather than microglia show a stronger binding affinity to neurons than microglia. In some embodiments, fusion proteins or fusion protein dimers that preferentially target neurons bind to neurons with an affinity at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 25 times that of microglia.

[0382] IGBFPL1 peptides or IGBFPL1 fragments that exhibit "preferential targeting" of microglia rather than neurons show a stronger binding affinity to microglia than neurons. In some embodiments, IGBFPL1 peptides or IGBFPL1 fragments that preferentially target microglia bind to microglia with an affinity at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 25 times that of neurons.

[0383] Fusion proteins or fusion protein dimers that exhibit “preferential targeting” of microglia rather than neurons show a stronger binding affinity to microglia than neurons. In some embodiments, fusion proteins or fusion protein dimers that preferentially target microglia bind to microglia with an affinity at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 25 times greater than that of neurons.

[0384] Therefore, this document provides a method for preferentially targeting neurons rather than microglia in a sample or subject, comprising introducing an IGBBPL1 fragment containing the C-terminal domain of IGBBPL1 into the sample or subject. This document also provides a method for preferentially targeting neurons rather than microglia in a sample or subject, comprising introducing an N-terminal fusion protein dimer as described herein into the sample or subject. In some embodiments, fusion proteins that preferentially bind to DCC relative to SORT1 are used to preferentially target neurons rather than microglia.

[0385] Therefore, this paper provides a method for preferentially targeting microglia rather than neurons in a sample or subject, comprising introducing an IGBBPL1 fragment containing the N-terminal domain of IGBBPL1 into the sample or subject. This paper also provides a method for preferentially targeting microglia rather than neurons in a sample or subject, comprising introducing a C-terminal fusion protein dimer as described herein into the sample or subject.

[0386] Treatment

[0387] In some respects, this disclosure provides methods for treating subjects with a disease or condition using the IGBBPL1 peptide, IGBBPL1 fragment, fusion protein, or composition described herein.

[0388] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject the IGBBPL1 peptide or a combination thereof described herein. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject the IGBBPL1 fragment or a combination thereof described herein. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject the fusion protein or a combination thereof described herein.

[0389] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject the IGBBPL1 peptide or a composition thereof described herein, wherein the IGBBPL1 peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 151-157. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject the IGBBPL1 peptide or a composition thereof described herein, wherein the IGBBPL1 peptide comprises an amino acid sequence selected from SEQ ID No: 151-157.

[0390] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179.

[0391] In some aspects, this disclosure provides a method for treating a subject with retinal degenerative disease, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179, provided that other domains of the IGBBPL1 polypeptide are not present in the IGBBPL1 fragment. In some aspects, this disclosure provides a method for treating a subject with retinal degenerative disease, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179, provided that other domains of the IGBBPL1 polypeptide are not present in the IGBBPL1 fragment.

[0392] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID No: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184.

[0393] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 15. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 15.

[0394] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 16. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 16.

[0395] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 17. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 17.

[0396] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 18. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 18.

[0397] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 19. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 19.

[0398] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 20. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 20.

[0399] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 21. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 21.

[0400] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 22. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 22.

[0401] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 51. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 51.

[0402] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 52. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 53.

[0403] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 54. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 54.

[0404] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 55. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 55.

[0405] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 56. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 56.

[0406] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 57. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 57.

[0407] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 58. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 58.

[0408] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 59. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 59.

[0409] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 60. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 60.

[0410] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 61. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 61.

[0411] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 62. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 62.

[0412] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 63. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 63.

[0413] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 64. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 64.

[0414] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 65. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 65.

[0415] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 66. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 66.

[0416] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 67. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 67.

[0417] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 68. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 68.

[0418] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 69. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 69.

[0419] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 70. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 70.

[0420] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 71. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 71.

[0421] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 72. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 72.

[0422] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 73. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 73.

[0423] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 74. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 74.

[0424] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 75. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 75.

[0425] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 76. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 76.

[0426] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 77. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 77.

[0427] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 78. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 78.

[0428] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 79. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 79.

[0429] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 80. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 80.

[0430] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 81. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 81.

[0431] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 82. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 82.

[0432] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 83. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 83.

[0433] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 84. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 84.

[0434] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 85. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 85.

[0435] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 86. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 86.

[0436] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 94. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 94.

[0437] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 95. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 95.

[0438] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 96. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 96.

[0439] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 97. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 97.

[0440] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 98. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 98.

[0441] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 99. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 99.

[0442] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 100. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 100.

[0443] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 101. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 101.

[0444] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 102. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 102.

[0445] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 103. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 103.

[0446] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 104. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 104.

[0447] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 105. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 105.

[0448] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 106. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 106.

[0449] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 107. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 107.

[0450] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 108. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 108.

[0451] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 109. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 109.

[0452] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 110. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 110.

[0453] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 111. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 111.

[0454] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 112. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 112.

[0455] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 113. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 113.

[0456] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 114. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 114.

[0457] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 115. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 115.

[0458] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 116. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 116.

[0459] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 117. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 117.

[0460] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 118. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 118.

[0461] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 119. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 119.

[0462] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 120. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 120.

[0463] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 121. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 121.

[0464] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 122. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 122.

[0465] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 123. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 123.

[0466] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 124. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 124.

[0467] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 125. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 125.

[0468] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 126. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 126.

[0469] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 127. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 127.

[0470] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 128. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 128.

[0471] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 129. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 129.

[0472] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 130. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 130.

[0473] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 131. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 131.

[0474] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 132. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 132.

[0475] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 133. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 133.

[0476] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 134. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 134.

[0477] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 135. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 135.

[0478] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 136. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 136.

[0479] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 138. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 138.

[0480] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 139. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 139.

[0481] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 140. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 140.

[0482] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 142. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 142.

[0483] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 144. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 144.

[0484] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 145. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 145.

[0485] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 146. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 146.

[0486] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 147. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 147.

[0487] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 148. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 148.

[0488] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 149. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 149.

[0489] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 150. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 150.

[0490] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 183. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 183.

[0491] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 184. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 184.

[0492] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 20, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 21. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 20, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 21.

[0493] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 22, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 9. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 22, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 9.

[0494] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 134, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 21. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 134, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 21.

[0495] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 138, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 139. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 138, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 139.

[0496] In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 140, and (ii) a second polypeptide comprising an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence shown in SEQ ID NO: 141. In some aspects, this disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein dimer comprising (i) a first polypeptide comprising the amino acid sequence of SEQ ID NO: 140, and (ii) a second polypeptide comprising the amino acid sequence of SEQ ID NO: 141.

[0497] In some aspects, this disclosure provides a method for treating a subject suffering from a retinal degenerative disease, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence. In some embodiments, the retinal degenerative disease is selected from glaucoma, dry age-related macular degeneration (AMD), geographic atrophy, wet AMD, retinitis pigmentosa, diabetic retinopathy, and Stagger's disease. In some embodiments, the retinal degenerative disease is glaucoma. In some embodiments, the retinal degenerative disease is dry AMD. In some embodiments, the retinal degenerative disease is wet AMD. In some embodiments, the retinal degenerative disease is retinitis pigmentosa. In some embodiments, the retinal degenerative disease is diabetic retinopathy. In some embodiments, the retinal degenerative disease is Stagger's disease.

[0498] In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject the IGBBPL1 peptide or a combination thereof described herein. In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject the IGBBPL1 fragment or a combination thereof described herein. In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject the fusion protein or a combination thereof described herein.

[0499] In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject the IGBBPL1 peptide or a composition thereof described herein, wherein the IGBBPL1 peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 151-157. In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject the IGBBPL1 peptide or a composition thereof described herein, wherein the IGBBPL1 peptide comprises an amino acid sequence selected from SEQ ID No: 151-157.

[0500] In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179. In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179.

[0501] In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179, provided that other domains of the IGBBPL1 polypeptide are not present in the IGBBPL1 fragment. In some aspects, this disclosure provides a method for treating a subject with a retinitis-related retinitis, comprising administering to the subject an IGBBPL1 fragment or composition described herein, wherein the IGBBPL1 fragment comprises an amino acid sequence selected from SEQ ID No: 2-3, 25-28, 37-50, 158, and 160-179, provided that other domains of the IGBBPL1 polypeptide are not present in the IGBBPL1 fragment.

[0502] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from SEQ ID No: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID No: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184.

[0503] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 15. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 15.

[0504] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 16. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 16.

[0505] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 17. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 17.

[0506] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 18. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 18.

[0507] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 19. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 19.

[0508] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 20. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 20.

[0509] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 21. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 21.

[0510] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 22. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 22.

[0511] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 51. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 51.

[0512] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 52. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 53.

[0513] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 54. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 54.

[0514] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 55. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 55.

[0515] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 56. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 56.

[0516] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 57. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 57.

[0517] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 58. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 58.

[0518] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 59. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 59.

[0519] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 60. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 60.

[0520] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 61. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 61.

[0521] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 62. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 62.

[0522] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 63. In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 63.

[0523] In some aspects, this disclosure provides a method for treating a subject with an inflammatory retinitis, comprising administering to the subject a fusion protein or composition described herein, wherein the fusion protein comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, ...

Claims

1. A fusion protein comprising a half-life extension domain and a human IGFBPL1 (hIGFBPL1) polypeptide or a fragment thereof, wherein the half-life extension domain is operatively linked to the hIGFBPL1 polypeptide or a fragment thereof with or without a linker.

2. The fusion protein of claim 1, wherein the hIGFBPL1 polypeptide or a fragment thereof comprises a thumb domain, an N-terminal domain, a Kazal domain and an Ig-like domain, a C-terminal domain, or any combination thereof.

3. The fusion protein of claim 2, wherein the thumb domain comprises the amino acid sequence of SEQ ID NO:25, the N-terminal domain comprises the amino acid sequence of SEQ ID NO:2, the Kazal domain comprises the amino acid sequence of SEQ ID NO:27, the Ig-like domain comprises the amino acid sequence of SEQ ID NO:26, and / or the C-terminal domain comprises the amino acid sequence of SEQ ID NO:

3.

4. The fusion protein according to any one of claims 1 to 3, wherein the fragment of hIGFBPL1 does not contain the thumb domain of hIGFBPL1.

5. The fusion protein according to any one of claims 1 to 4, wherein the fragment of hIGFBPL1 does not contain the N-terminal domain of hIGFBPL1.

6. The fusion protein according to any one of claims 1 to 5, wherein the fragment of hIGFBPL1 does not contain the Kazal domain of hIGFBPL1.

7. The fusion protein according to any one of claims 1 to 6, wherein the fragment of hIGFBPL1 does not contain the Ig-like domain of hIGFBPL1.

8. The fusion protein of any one of claims 1 to 7, wherein the fragment of hIGFBPL1 does not contain the C-terminal domain.

9. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the N-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

10. The fusion protein of claim 9, wherein the fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID NO:2, 158, and 160-166.

11. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

12. The fusion protein of claim 11, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:

179.

13. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the thumb domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

14. The fusion protein of claim 13, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:

25.

15. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

16. The fusion protein of claim 15, wherein the fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID No: 26, 167, and 169.

17. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the Kazal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

18. The fusion protein of claim 17, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:27 or SEQ ID NO:

178.

19. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the Ig-like domain and the C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

20. The fusion protein of claim 19, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:168 or SEQ ID NO:

170.

21. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the Kazal domain and the Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

22. The fusion protein of claim 21, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:171 or SEQ ID NO:

173.

23. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the Kazal domain, the Ig-like domain, and the C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

24. The fusion protein of claim 23, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:172 or SEQ ID NO:

174.

25. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the N-terminal domain and the Ig-like domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

26. The fusion protein of claim 25, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:

175.

27. The fusion protein of any one of claims 1 to 3, wherein the fragment of hIGFBPL1 comprises the N-terminal domain, the Ig-like domain, and the C-terminal domain of hIGFBPL1, provided that other domains of hIGFBPL1 are not part of the hIGFBPL1 fragment.

28. The fusion protein of claim 27, wherein the fragment of hIGFBPL1 comprises the amino acid sequence of SEQ ID NO:176 or SEQ ID NO:

177.

29. The fusion protein of any one of claims 1 to 24, wherein the hIGFBPL1 polypeptide or the fragment of hIGFBPL1 comprises one or more domains of mouse IGFBPL1 (mIGFBPL1).

30. The fusion protein of claim 29, wherein the hIGFBPL1 polypeptide or the fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID No: 151-153 and 155-157.

31. The fusion protein of any one of claims 1 to 30, wherein the hIGFBPL1 polypeptide or a fragment thereof comprises the natural signal peptide of hIGFBPL1.

32. The fusion protein of claim 31, wherein the natural signal peptide comprises the amino acid sequence of SEQ ID NO:

28.

33. The fusion protein of any one of claims 1 to 30, wherein the hIGFBPL1 polypeptide or a fragment thereof comprises a heterologous signal peptide.

34. The fusion protein of claim 33, wherein the heterologous signal peptide comprises the amino acid sequence of SEQ ID NO:

36.

35. The fusion protein of any one of claims 1 to 3 and 31 to 34, wherein the hIGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID NO: 1, 10 and 11, or wherein the fragment of hIGFBPL1 comprises an amino acid sequence selected from SEQ ID NO: 37-50, 158 and 160-180.

36. The fusion protein according to any one of claims 1 to 35, wherein the extended half-life domain is located at the N-terminus of the hIGFBPL1 polypeptide or a fragment thereof.

37. The fusion protein of any one of claims 1 to 35, wherein the extended half-life domain is located at the C-terminus of the hIGFBPL1 polypeptide or a fragment thereof.

38. The fusion protein according to any one of claims 1 to 37, wherein the extended half-life domain is selected from the Fc domain, albumin domain, transferrin domain, C-terminal peptide, elastin-like peptide, inaccurate repeating peptide sequence, proline-alanine-serine polymer, repeating sequence of glycine-rich polypeptide, and gelatin-like protein.

39. The fusion protein according to any one of claims 1 to 37, wherein the extended half-life domain is an Fc domain.

40. The fusion protein of claim 39, wherein the Fc domain comprises an amino acid sequence selected from SEQ ID NO: 4-9.

41. The fusion protein according to any one of claims 1 to 37, wherein the extended half-life domain is an albumin polypeptide.

42. The fusion protein of claim 41, wherein the albumin polypeptide comprises the amino acid sequence of SEQ ID NO:

181.

43. The fusion protein of any one of claims 1 to 42, wherein the half-life extension domain and the hIGFBPL1 polypeptide or a fragment thereof are operatively linked without a linker.

44. The fusion protein of any one of claims 1 to 42, wherein the half-life extension domain and the hIGFBPL1 polypeptide or a fragment thereof are operatively linked in the presence of a linker.

45. The fusion protein of claim 44, wherein the linker is a peptide linker.

46. ​​The fusion protein of claim 44 or 45, wherein the adapter is a TEV adapter or a GS adapter.

47. The fusion protein of any one of claims 44 to 46, wherein the linker comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NO: 12-14, 23-24, 159 and 182.

48. The fusion protein of claim 1, wherein the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO: 15-22, 109-110, 134, 138-140, 142, 148-150.

49. The fusion protein of claim 48, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 15-22, 109-110, 134, 138-140, 142, 148-150.

50. The fusion protein of claim 1, wherein the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO:51-72, 111-133, and 183.

51. The fusion protein of claim 50, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 51-72, 111-133 and 183.

52. The fusion protein of claim 1, wherein the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID NO: 94-100 and 102-108.

53. The fusion protein of claim 52, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 94-100 and 102-108.

54. The fusion protein of claim 1, wherein the fusion protein comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the amino acid sequence selected from SEQ ID No: 144-147.

55. The fusion protein of claim 54, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID NO: 144-147.

56. The fusion protein of any one of claims 1 to 55, wherein the hIGFBPL1 or a fragment thereof contains at least one glycosylation mutation.

57. The fusion protein of claim 56, wherein the glycosylation mutation is at amino acid residues N166 and / or S268 as identified in SEQ ID NO:

1.

58. The fusion protein of claim 57, wherein the glycosylation mutation is N166Q, N166D, S268A or any combination thereof.

59. The fusion protein of claim 57, wherein the hIGFBPL1 polypeptide comprising the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NO: 87-91.

60. The fusion protein of any one of claims 38 to 59, wherein the Fc domain comprises a glycosylation mutation, optionally wherein the glycosylation mutation of the Fc domain is at amino acid residue N297 as identified in any one of SEQ ID NO: 4-9.

61. The fusion protein of claim 60, wherein the glycosylation mutation of the Fc domain is N297D or N297Q.

62. The fusion protein of claim 60, wherein the Fc domain comprising a glycosylation mutation comprises an amino acid sequence selected from SEQ ID NO: 92 and 93.

63. A fusion protein dimer comprising two fusion proteins as described in any one of claims 1 to 62.

64. The fusion protein dimer of claim 63, wherein the two fusion proteins are identical.

65. The fusion protein dimer of claim 63, wherein the two fusion proteins are different.

66. A fusion protein dimer comprising (i) a first fusion protein comprising a first half-life extension domain operatively linked to a first hIGFBPL1 polypeptide or a fragment thereof with or without a linker, and (ii) a second fusion protein comprising a second half-life extension domain operatively linked to a second hIGFBPL1 polypeptide or a fragment thereof with or without a linker.

67. The fusion protein dimer of claim 66, wherein the first and second IGBBPL1 polypeptides or fragments thereof are identical.

68. The fusion protein dimer of claim 66, wherein the first and second IGBBPL1 polypeptides or fragments thereof are different.

69. The fusion protein dimer of any one of claims 66 to 68, wherein the first and second half-life extension domains are identical.

70. The fusion protein dimer of any one of claims 66 to 68, wherein the first and second half-life extension domains are distinct.

71. The fusion protein dimer of any one of claims 66 to 70, wherein the first half-life extension domain is operatively linked to the N-terminus of the first hIGFBPL1 polypeptide or a fragment thereof.

72. The fusion protein dimer of any one of claims 66 to 70, wherein the first half-life extension domain is operatively linked to the C-terminus of the first hIGFBPL1 polypeptide or a fragment thereof.

73. The fusion protein dimer of any one of claims 66 to 72, wherein the second half-life extension domain is operatively linked to the N-terminus of the second hIGFBPL1 polypeptide or a fragment thereof.

74. The fusion protein dimer of any one of claims 66 to 72, wherein the second half-life extension domain is operatively linked to the C-terminus of the second hIGFBPL1 polypeptide or a fragment thereof.

75. The fusion protein dimer of any one of claims 66 to 74, wherein the first fusion protein comprises a third hIGFBPL1 polypeptide or a fragment thereof operatively linked to the half-life extension domain with or without a linker.

76. The fusion protein dimer of claim 75, wherein the first and third hIGFBPL1 polypeptides or fragments thereof are identical.

77. The fusion protein dimer of claim 75, wherein the first and third hIGFBPL1 polypeptides or fragments thereof are different.

78. The fusion protein dimer of any one of claims 75 to 77, wherein the third hIGFBPL1 polypeptide or fragment thereof is operatively linked to the opposite end of the half-life extension domain compared to the first hIGFBPL1 polypeptide or fragment thereof.

79. The fusion protein dimer of any one of claims 66 to 78, wherein the second fusion protein comprises a fourth hIGFBPL1 polypeptide or a fragment thereof operatively linked to the half-life extension domain with or without a linker.

80. The fusion protein dimer of claim 79, wherein the second and fourth hIGFBPL1 polypeptides or fragments thereof are identical.

81. The fusion protein dimer of claim 79, wherein the second and fourth hIGFBPL1 polypeptides or fragments thereof are different.

82. The fusion protein dimer of any one of claims 79 to 81, wherein the fourth hIGFBPL1 polypeptide or fragment thereof is operatively linked to the opposite end of the half-life extension domain compared to the second hIGFBPL1 polypeptide or fragment thereof.

83. The fusion protein dimer of any one of claims 66 to 82, wherein the first, second, third and / or fourth hIGFBPL1 polypeptide or fragment thereof comprises an amino acid sequence selected from SEQ ID NO:1, 10-11, 37-50, 158 and 160-180.

84. The fusion protein dimer of any one of claims 66 to 83, wherein the first half-life extension domain is a first Fc domain, and the second half-life extension domain is a second Fc domain.

85. The fusion protein dimer of claim 84, wherein the first and second Fc domains contain mutations for dimerization.

86. The fusion protein dimer of claim 85, wherein the first Fc domain comprises a club-shaped mutation and the second Fc domain comprises a mortar-shaped mutation to dimerize the first and second Fc domains.

87. The fusion protein dimer of claim 85, wherein the first Fc domain comprises a mortar mutation and the second Fc domain comprises a pestle mutation to dimerize the first and second Fc chains.

88. The fusion protein dimer of any one of claims 84 to 86, wherein the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NO:4-9, 92 and 93.

89. The fusion protein dimer according to any one of claims 66 to 88, wherein: (i) the first fusion protein comprises the amino acid sequence of SEQ ID NO:16, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:16; (ii) the first fusion protein comprises the amino acid sequence of SEQ ID NO:15, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:15; (iii) the first fusion protein comprises the amino acid sequence of SEQ ID NO:19, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:19; (iv) the first fusion protein comprises the amino acid sequence of SEQ ID NO:20, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:21; (v) the first fusion protein comprises the amino acid sequence of SEQ ID NO:22, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:9; (vi) the first fusion protein comprises the amino acid sequence of SEQ ID NO:134, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:21; (vii) the first fusion protein comprises the amino acid sequence of SEQ ID NO:138, and the second fusion protein comprises the amino acid sequence of SEQ ID NO:139; or (viii) the first fusion protein comprises the amino acid sequence of SEQ ID NO:

15. The second fusion protein contains the amino acid sequence of SEQ ID NO: 140, and the second fusion protein contains the amino acid sequence of SEQ ID NO:

141.

90. A fusion protein dimer comprising the fusion protein as described in any one of claims 1 to 62 and a second half-life extension domain, wherein the second half-life extension domain does not contain the hIGFBPL1 polypeptide or a fragment thereof.

91. The fusion protein dimer of claim 90, wherein the first and second half-life extension domains are identical.

92. The fusion protein dimer of claim 90, wherein the first and second half-life extension domains are different.

93. The fusion protein dimer of any one of claims 90 to 92, wherein the first half-life extension domain is a first Fc domain, and the second half-life extension domain is a second Fc domain.

94. The fusion protein dimer of claim 93, wherein the first Fc domain comprises a club-shaped mutation and the second Fc domain comprises a mortar-shaped mutation to dimerize the first and second Fc domains.

95. The fusion protein dimer of claim 93, wherein the first Fc domain comprises a mortar mutation and the second Fc domain comprises a pestle mutation to dimerize the first and second Fc domains.

96. The fusion protein dimer of any one of claims 93 to 95, wherein the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NO:4-9, 92, and 93.

97. The fusion protein dimer of any one of claims 90 to 96, wherein the first fusion protein comprises a second hIGFBPL1 polypeptide or a fragment thereof operatively linked to the first half-life extension domain with or without a linker.

98. The fusion protein dimer of claim 97, wherein the second hIGFBPL1 polypeptide or a fragment thereof is operatively linked to the opposite end of the hIGFBPL1 polypeptide or a fragment thereof.

99. The fusion protein dimer of claim 97 or 98, wherein the first fusion protein comprises the amino acid sequence of SEQ ID NO:

22.

100. A fusion protein dimer comprising a first fusion protein and a second fusion protein, wherein each of the first and second fusion proteins comprises an amino acid sequence selected from SEQ ID NO:51-86, wherein the first and second fusion proteins are the same or different.

101. A composition comprising a fusion protein or a fusion protein dimer as described in any one of claims 1 to 100.

102. A method for inhibiting DCC activity in a sample or subject, comprising introducing a fusion protein or fusion protein dimer into the sample or subject as described in any one of claims 1 to 3, 7 to 10, 25 to 34 and 37 to 100.

103. The method of claim 102, wherein the fusion protein or fusion protein dimer binds to DCC in the cell.

104. The method of claim 103, wherein the cell is a neuron.

105. The method of claim 104, wherein the fusion protein or fusion protein dimer increases the average neurite length, the number of neurites, the number of neurons, and / or the number of neurites per neuron.

106. The method of claim 104 or 105, wherein the fusion protein increases neurotrophic factor signaling.

107. A method for inhibiting SORT1 activity in a sample or subject, comprising introducing a fusion protein or fusion protein dimer into the sample or subject as described in any one of claims 1 to 5, 15 to 16, 19 to 24, 31 to 34, 36, and 38 to 100.

108. The method of claim 107, wherein the fusion protein or fusion protein dimer binds SORT1 in the cell.

109. The method of claim 108, wherein the cell is a microglia.

110. The method of claim 109, wherein the fusion protein or fusion protein dimer increases the secretion of at least one cytokine by the microglia.

111. The method of claim 110, wherein the at least one cytokine is selected from IL-10, GM-CSF, MIP-1α, and any combination thereof.

112. The method of any one of claims 109 to 111, wherein the fusion protein or fusion protein dimer inhibits the microglia from producing one or more proteins.

113. The method of claim 112, wherein the one or more proteins comprise one or more inflammatory cytokines.

114. The method of claim 113, wherein the one or more inflammatory cytokines comprise IL-6.

115. The method of any one of claims 112 to 114, wherein the one or more proteins comprise IP-10.

116. The method of any one of claims 109 to 115, wherein the fusion protein or fusion protein dimer increases the phagocytic activity of the microglia.

117. The method of any one of claims 108 to 116, wherein the cell is a neuron.

118. The method of claim 117, wherein the fusion protein or fusion protein dimer increases the average neurite length, the number of neurites, the number of neurons, and / or the number of neurites per neuron.

119. The method of claim 117 or 118, wherein the fusion protein increases neurotrophic factor signaling.

120. A method of treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or a fusion protein dimer as described in any one of claims 1 to 100 or a composition as described in claim 101.

121. The method of claim 120, wherein the retinal degenerative disease is glaucoma, dry age-related macular degeneration (AMD), geographic atrophy, wet AMD, retinitis pigmentosa, diabetic retinopathy, or Staggart's disease.

122. A method of treating a subject with inflammatory retinal disease, comprising administering to the subject a fusion protein or fusion protein dimer as described in any one of claims 1 to 100 or a composition as described in claim 101.

123. The method of claim 122, wherein the retinal inflammatory disease is retinal vasculitis, uveitis, NAION, or neuromyelitis optica.

124. The method of any one of claims 120 to 123, wherein the fusion protein, fusion protein dimer, or composition is applied to the eye of the subject.

125. The method of claim 124, wherein application to the eye comprises epidermal application, topical ocular application, or systemic application.

126. The method of claim 125, wherein topical ocular application includes periocular, intravitreal, or intraanterior chamber application.

127. A method of treating a subject with a neurodegenerative disease, comprising administering to the subject a fusion protein or fusion protein dimer as described in any one of claims 1 to 100 or a composition as described in claim 101.

128. The method of claim 127, wherein the fusion protein, fusion protein dimer, or composition is administered parenterally to the subject.

129. The method of claim 128, wherein the fusion protein, fusion protein dimer, or composition is administered subcutaneously or intravenously to the subject.

130. The method of any one of claims 127 to 129, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, frontotemporal dementia, amyotrophic lateral sclerosis, progressive supranuclear palsy, leukoencephalopathy, multiple sclerosis, transverse myelitis, and Sheldr's disease.

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