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

Abstract

This 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] INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN LIKE 1 (IGFBPL1) COMPOSITIONS AND METHODS OF USE THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 510,577, filed June 27, 2023, which is incorporated by reference herein in its entirety.

[0004] BACKGROUND

[0005] Insulin-like growth factor binding protein like 1 (IGFBPL1) is a multi-domain protein reported to have beneficial activity in neurodegenerative and neuroinflammatory conditions such as glaucoma by, for example, promoting neurite outgrowth. However, the underlying mechanisms and activities resulting in IGFBPL1 activity in such conditions are unknown.

[0006] SUMMARY

[0007] In some aspects, the disclosure provides a fusion protein comprising a half-life extending domain and a human IGFBPL1 (hIGFBPLI) polypeptide or a fragment thereof, wherein the half- life extending domain is operatively linked to the hIGFBPLI polypeptide or fragment thereof with or without a linker.

[0008] In some aspects, the disclosure provides a fusion protein comprising a half-life extending domain and a means for binding DCC, wherein the half-life extending domain is operatively linked to the means with or without a linker.

[0009] In some aspects, the disclosure provides a fusion protein comprising a half-life extending domain and a means for binding SORT1, wherein the half-life extending domain is operatively linked to the means with or without a linker.

[0010] In some aspects, the disclosure provides a fusion protein comprising a half-life extending domain and a means for binding SORT1 and DCC, wherein the half-life extending domain is operatively linked to the means with or without a linker.

[0011] In some embodiments, the hIGFBPLI polypeptide or the fragment thereof comprises a thumb domain, an N-terminal domain, a Kazal domain, and 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.

[0012] In some embodiments, the fragment of hIGFBPLI does not comprise the thumb domain of hIGFBPLI. In some embodiments, the fragment of hIGFBPLI does not comprise the N- terminal domain of hIGFBPLI. In some embodiments, the fragment of hIGFBPLI does not comprise the Kazal domain of hIGFBPLI. In some embodiments, the fragment of hIGFBPLI does not comprise the Ig-like domain of hIGFBPLI. In some embodiments, the fragment of hIGFBPLI does not comprise the C-terminal domain.

[0013] In some embodiments, the fragment of hIGFBPLI comprises the N-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence selected from SEQ ID NOs: 2, 158, and 160-166.

[0014] In some embodiments, the fragment of hIGFBPLI comprises the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 179.

[0015] In some embodiments, the fragment of hIGFBPLI comprises the thumb domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 25.

[0016] In some embodiments, the fragment of hIGFBPLI comprises the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence selected from SEQ ID Nos: 26, 167, and 169.

[0017] In some embodiments, the fragment of hIGFBPLI comprises the Kazal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 27 or SEQ ID NO: 178.

[0018] In some embodiments, the fragment of hIGFBPLI comprises the Ig-like domain and C- terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 168 or SEQ ID NO: 170.

[0019] In some embodiments, the fragment of hIGFBPLI comprises the Kazal domain and the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 171 or SEQ ID NO: 173.

[0020] In some embodiments, the fragment of hIGFBPLI comprises the Kazal domain, the Ig- like domain, and the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 172 or SEQ ID NO: 174.

[0021] In some embodiments, the fragment of hIGFBPLI comprises the N-terminal domain and the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 175.

[0022] In some embodiments, the fragment of hIGFBPLI comprises the N-terminal domain, the Ig-like domain, and the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment. In some embodiments, the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 176 or SEQ ID NO: 177.

[0023] In some embodiments, the hIGFBPLI polypeptide or the fragment of hIGFBPLI comprises one or more domains of murine IGFBPL1 (mIGFBPLI). In some embodiments, the hIGFBPLI polypeptide or the fragment of hIGFBPLI comprises an amino acid sequence selected from SEQ ID Nos: 151-153 and 155-157.

[0024] In some embodiments, the hIGFBPLI polypeptide or the fragment thereof comprises the native signal peptide of hIGFBPLI. In some embodiments, the native signal peptide comprises the amino acid sequence of SEQ ID NO: 28. In some embodiments, the hIGFBPLI polypeptide or the fragment thereof comprises a heterologous signal peptide. In some embodiments, the heterologous signal peptide comprises the amino acid sequence of SEQ ID NO: 36.

[0025] In some embodiments, the hIGFBPLI polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 1, 10 and 11, or the fragment of hIGFBPLI comprises an amino acid sequence selected from SEQ ID NOs: 37-50. In some embodiments, the hIGFBPLI polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 1, 10 and 11, or wherein the fragment of hIGFBPLI comprises an amino acid sequence selected from SEQ ID NOs: 37-50, 158 and 160-180.

[0026] In some embodiments, the half-life extending domain is located at the N-terminus of the hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the half-life extending domain is located at the C-terminus of the hIGFBPLI polypeptide or the fragment thereof.

[0027] In some embodiments, the half-life extending domain is selected from an Fc domain, an albumin domain, a transferrin domain, a C-terminal peptide, an elastin-like peptide, a non-exact repeat peptide sequence, a proline-alanine-serine polymer, a repeat sequence of glycine rich polypeptides, and a gelatin-like protein.

[0028] In some embodiments, the half-life extending domain is an Fc domain. In some embodiments, the Fc domain comprises an amino acid sequence selected from SEQ ID NOs: 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.

[0029] In some embodiments, the half-life extending domain is an albumin polypeptide. In some embodiments, the albumin polypeptide comprises the amino acid sequence of SEQ ID NO: 181.

[0030] In some embodiments, the half-life extending domain and the hIGFBPLI polypeptide or the fragment thereof are operably linked without a linker. In some embodiments, the half-life extending domain and the hIGFBPLI polypeptide or the 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 NOs: 12-14. In some embodiments, the linker comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 12-14, 23-24, 159, and 182.

[0031] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 15-22, 109- 110, 134, 138-140, 142, 148-150. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 15-22, 109-110, 134, 138-140, 142, 148-150.

[0032] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 51-72, 111- 133 and 183. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 51-72, 111-133 and 183.

[0033] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 94-100, and 102-108. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 94-100, and 102-108.

[0034] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID Nos: 144-147. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 144-147.

[0035] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0036] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0037] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0038] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0039] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0040] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0041] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0042] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to 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.

[0043] In some embodiments, the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 51-72. In some embodiments, the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 51-72.

[0044] In some embodiments, the hIGFBPLI or the fragment thereof comprises at least one glycosylation mutation. In some embodiments, the glycosylation mutation is at amino acid residue 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 hIGFBPLI polypeptide comprising the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NOs: 87-91.

[0045] In some embodiments, the Fc domain comprises a glycosylation mutation. In some embodiments, the glycosylation mutation of the Fc domain is at amino acid residue N297 as identified in any one of SEQ ID NOs: 4-9. In some embodiments, the glycosylation mutation of the Fc domain is N297D or N297Q. In some embodiments, the Fc domain comprising the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NOs: 92 and 93.

[0046] In some aspects, the disclosure provides a fusion protein dimer comprising two fusion proteins of those disclosed herein. In some embodiments, the two fusion proteins are the same. In some embodiments, the two fusion proteins are different.

[0047] In some aspects, the disclosure provides a fusion protein dimer comprising (i) a first fusion protein comprising a first half-life extending domain operably linked to a first hIGFBPLI polypeptide or a fragment thereof with or without a linker, and (ii) a second fusion protein comprising a second half-life extending domain operably linked to a second hIGFBPLI polypeptide or a fragment thereof with or without a linker.

[0048] In some embodiments, the first and second IGFBPL1 polypeptide or the fragment thereof is the same. In some embodiments, the first and second IGFBPL1 polypeptide or the fragment thereof is different.

[0049] In some embodiments, the first and second half-life extending domain is the same. In some embodiments, the first and second half-life extending domain is different. In some embodiments, the first half-life extending domain is operably linked to the N-terminus of the first hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the first half-life extending domain is operably linked to the C-terminus of the first hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the second half-life extending domain is operably linked to the N-terminus of the second hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the second half-life extending domain is operably linked to the C-terminus of the second hIGFBPLI polypeptide or the fragment thereof.

[0050] In some embodiments, the first fusion protein comprises a third hIGFBPLI polypeptide or a fragment thereof operably linked to the half-life extending domain with or without a linker. In some embodiments, the first and third hIGFBPLI polypeptides or the fragments thereof are the same. In some embodiments, the first and third hIGFBPLI polypeptides or the fragments thereof are different. In some embodiments, the third hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the half-life extending domain than the first hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the second fusion protein comprises a fourth hIGFBPLI polypeptide or a fragment thereof operably linked to the half-life extending domain with or without a linker. In some embodiments, the second and fourth hIGFBPLI polypeptides or the fragments thereof are the same. In some embodiments, the second and fourth hIGFBPLI polypeptides or the fragments thereof are different. In some embodiments, the fourth hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the half-life extending domain than the second hIGFBPLI polypeptide or the fragment thereof.

[0051] In some embodiments, the first, second, third and / or fourth hIGFBPLI polypeptide or the fragment thereof, comprises an amino acid sequence selected from SEQ ID NOs: 1, 10-11, and 37-50. In some embodiments, the first, second, third and / or fourth hIGFBPLI polypeptide or the fragment thereof, comprises an amino acid sequence selected from SEQ ID NOs: 1, 10-11, 37- 50, 158 and 160-180.

[0052] In some embodiments, the first half-life extending domain is a first Fc domain and the second half-life extending domain is a second Fc domain. In some embodiments, the first and second Fc domains comprise mutations for dimerization. In some embodiments, the first Fc domain comprises knob mutations and the second Fc domain comprises hole mutations to dimerize the first and second Fc domains. In some embodiments, the first Fc domain comprises hole mutations and the second Fc domain comprises knob mutations to dimerize the first and second Fc chains. In some embodiments, the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NOs: 4-9, 92 and 93.

[0053] 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, wherein 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.

[0054] In some aspects, the disclosure provides a fusion protein dimer comprising a fusion protein described herein, and a second half-life extending domain, wherein the second half-life extending domain does not comprise an hIGFBPLI polypeptide or a fragment thereof. In some embodiments, the first and second half-life extending domain is the same. In some embodiments, the first and second half-life extending domain is different. In some embodiments, the first halflife extending domain is a first Fc domain and the second half-life extending domain is a second Fc domain. In some embodiments, the first Fc domain comprises knob mutations and the second Fc domain comprises hole mutations to dimerize the first and second Fc domains. In some embodiments, the first Fc domain comprises hole mutations and the second Fc domain comprises knob mutations 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 NOs: 4-9, 92 and 93. In some embodiments, the first fusion protein comprises a second hIGFBPLI polypeptide or a fragment thereof operably linked to the first half-life extending domain with or without a linker. In some embodiments, the second hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the hIGFBPLI polypeptide or the fragment thereof. In some embodiments, the first fusion protein comprises the amino acid sequence of SEQ ID NO: 22.

[0055] In some aspects, the 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 NOs: 51-86, wherein the first and second fusion proteins are the same or different.

[0056] In some aspects, the disclosure provides a composition comprising the fusion protein or fusion protein dimer disclosed herein.

[0057] In some aspects, the disclosure provides a method for inhibiting DCC activity in a sample or a subject, comprising introducing to the sample or the subject the fusion protein or fusion protein dimer disclosed herein. In some embodiments, the fusion protein or fusion protein dimer binds DCC in a cell. In some embodiments, the cell is a neuron. In some embodiments, the fusion protein or fusion protein dimer increases average neurite length, number of neurites, number of neurons, and / or number of neurites per neuron. In some embodiments, the fusion protein increases neurotrophin signaling.

[0058] In some aspects, the disclosure provides a method for inhibiting SORT1 activity in a sample or a subject, comprising introducing to the sample or the subject the fusion protein or fusion protein dimer disclosed herein. In some embodiments, the fusion protein or fusion protein dimer binds SORT1 in a cell. In some embodiments, the cell is a microglia. In some embodiments, the fusion protein or fusion protein dimer increases secretion of at least one cytokine by the microglia. In some embodiments, the at least one cytokine is selected from IL- 10, GM-CSF, MIP-1 alpha, and any combination thereof. In some embodiments, the at least one cytokine is selected from IL- 10, GM-CSF, MIP-1 alpha, CXCL8 (IL-8) and any combination thereof. In some embodiments, the fusion protein or fusion protein dimer inhibits production of one or more proteins by the 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 phagocytic activity of the microglia. In some embodiments, the cell is a neuron. In some embodiments, the fusion protein or fusion protein dimer increases average neurite length, number of neurites, number of neurons, and / or number of neurites per neuron. In some embodiments, the fusion protein increases neurotrophin signaling.

[0059] In some aspects, the disclosure provides a method for preferential targeting of neurons over microglia in a sample or subject, comprising introducing to the sample or subject the fusion protein or the fusion protein dimer described herein.

[0060] In some aspects, the disclosure provides a method for preferential targeting of neurons over microglia in a sample or subject, comprising introducing to the sample or subject a fusion protein comprising a means for binding DCC and a half-life extending domain.

[0061] In some aspects, the disclosure provides a method of treating a retinal degenerative disease in a subject, comprising administering to the subject a fusion protein, a fusion protein dimer, or a 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.

[0062] In some aspects, the disclosure provides a method of treating a retinal inflammatory disease in a subject, comprising administering to the subject a fusion protein, a fusion protein dimer, or a composition described herein. In some embodiments, the retinal inflammatory disease is retinal vasculitis, uveitis, NAION, or neuromyelitis optica.

[0063] In some embodiments, the fusion protein, fusion protein dimer, or composition is administered to the eye of the subject. In some embodiments, administration to the eye comprises topical administration, local ocular administration, or systemic administration. In some embodiments, local ocular administration comprises periocular, intravitreal, or intracameral administration.

[0064] In some aspects, the disclosure provides a method of treating a neurodegenerative disease in a subject, comprising administering to the subject a fusion protein, a fusion protein dimer, or a composition described herein. In some embodiments, the fusion protein, fusion protein dimer, or composition is parenterally administered to the subject. In some embodiments, the fusion protein, fusion protein dimer, or composition is subcutaneously or intravenously administered 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.

[0065] BRIEF DESCRIPTION OF THE DRAWINGS

[0066] FIG. 1 provides a schematic showing combination of human IGBFPL1 protein domains of exemplary human IGFBPL1 (hIGFBPLI) constructs.

[0067] FIG. 2A provides schematics of the following hIGFBPLI constructs: PRO802 (bivalent IGFBPL1 N-terminus linked to Fc); PRO801 (bivalent IGFBPL1 C-terminus linked to Fc; PRO 1399 (monovalent IGFBPL1 C-terminus linked to Fc); and PRO 1400 (monovalent IGFBPL1 N-terminus linked to Fc).

[0068] FIGs. 2B and 2C provide graphs showing binding of the hIGFBPLI constructs of FIG.

[0069] 2A to MCF7 cells. FIG. 3 provides images showing binding of PRO801 at 200 nM concentration to cell bodies and a subset of synapses in hIPSC-derived neuron cultures.

[0070] FIG. 4 provides images showing binding of the hIGFBPLI constructs of FIG. 2A at 200 nM concentration to somata, neurites and synapses.

[0071] FIG. 5 provides images showing binding of the hIGFBPLI constructs of FIG. 2A at 200 nM concentration to microglia.

[0072] FIG. 6A provides images showing binding of PRO802 to iPSC-derived neurons.

[0073] FIGs. 6B-6E provides graphs quantifying neurite length (FIG. 6B), number of neurites (FIG. 6C), number of neurons (FIG. 6D), and number of neurites per neuron (FIG. 6E) of iPSC-derived neurons after culture with PRO802 and PRO801. Significance determined via post-hoc Dunnett’ s test (*p < 0.05, **p < 0.01, ***p < 0.001). All datasets exhibit significance by 1-way ANOVA. Data are from 3 fields of view each from 5 cultures.

[0074] FIGs. 7A-7C provide graphs showing the effect of PRO801 and PRO802 on cytokine secretion from hIPSC-derived microglia.

[0075] FIG. 8 provides graphs showing the binding of PRO801 and PRO802 to SORT1 and DCC as determined by Octet.

[0076] FIGs. 9A and 9B provide flow cytometry histograms of the constructs shown in FIG. 2A and assay controls, respectively.

[0077] FIG. 10 provides schematics of the following hIGFBPLI constructs: PRO1571 (tetravalent IGFBPL1 linked to Fc); PRO1572 (trans bivalent IGFBPL1 linked to Fc); and PRO1573 (cis bivalent IGFBPL1 linked to Fc).

[0078] FIGs. 11A-11C provides images showing binding of PRO801 and the constructs in FIG. 10 to microglia at concentrations of 200 nM (FIG. 11A) or 20 nM (FIG. 11B). FIG. 11C shows binding to neurons at 200 nM or 20 nM. Top panels show labeling of IGFBPL1 constructs and bottom panel shows labeling for microglia based on IBA-1 staining.

[0079] FIG. 12 provides a graph showing binding of PRO801 glycosylated (013) or deglycosylated (012) via PNGase to BeWo cells at varying concentrations.

[0080] FIG. 13A provides images showing binding of PRO801 glycosylated (013) or deglycosylated (012) to neurons at 200 nM or 20 nM.

[0081] FIG. 13B provides images showing binding of PRO801 glycosylated (013) or deglycosylated (012) to microglia at 20 nM. FIG. 14 provides a graph showing cell binding to BeWo cells (epithelial cells) of IGFBPLl-Fc fusion proteins having the indicated glycosylation mutations.

[0082] FIG. 15 provides graphs showing binding of a recombinant murine IGFBPL1 (PRO806) to human and mouse SORT1 and DCC via biolayer interferometry (BLI).

[0083] FIG. 16 provides a graph showing binding of IGFBPLl-Fc fusion proteins to microglia.

[0084] FIG. 17 provides a graph showing correlation of IGFBPLl-Fc fusion protein binding to microglia and SORT1.

[0085] FIG. 18 provides graphs showing effects of IGFBPLl-Fc fusion proteins on microglial phagocytosis.

[0086] FIGs. 19A-19B provide graphs showing suppression of expression of IL-6 (FIG. 19A) and IP-10 (FIG. 19B) protein in microglia cells treated with LPS by IGFBPLl-Fc fusion protein PRO1643 at indicated concentrations.

[0087] FIGs. 20A-20B provides graphs showing binding of IGFBPLl-Fc fusion proteins to neurons at a concentration of 20nM (FIG. 20A) or 200nM (FIG. 20B).

[0088] FIG. 21 provides a graph showing AKT activation in neurons by IGFBPLl-Fc fusion protein PRO 1643 at indicated concentrations.

[0089] FIG. 22 provides a graph showing neuron survival in cultures treated with IGFBPLl-Fc fusion PRO 1643 at indicated concentrations.

[0090] DETAILED DESCRIPTION

[0091] IGBFPL1 was previously described as binding to neurons and microglia (see, e.g., US Patent No. 10,842,849, and US Publication No. 2022-01431319, each hereby incorporated by this reference). However, all of the binding partners of IGFBPL1 were not known. The disclosure is based, at least in part, on the discovery that IGFBPL1 does not bind to IGF1R as previously predicted, but instead binds sortilin 1 (SORT1) in addition to the netrin 1 receptor (DCC). Specifically, as demonstrated herein, fusion proteins presenting the N-terminus of IGFBPL1 (i.e., fused to the C-terminus) were identified as only binding to DCC, whereas fusion proteins presenting the C-terminus of IGFBPL1 (i.e., fused to the N-terminus) were identified as binding to both DCC and SORT1. In addition, it was determined that the N-terminal domains of IGFBPL1 were required for binding to DCC, whereas the Ig-like domain was required for binding to SORT1. SORT1 is highly expressed in the brain and eye, particularly on microglia. S0RT1 is a major contributor to clearance of progranulin (PGRN) which blocks pro- inflammatory signaling and primes microglia for inflammation resolution. Without wishing to be bound by theory, IGFBPL1 provides anti-inflammatory and inflammation resolution due to inhibition of SORT1. Further, SORT1 functions as an ApoE receptor which is a known activator of microglia. Accordingly, it is believed based on the data provided herein that IGFBPL1 is competitive with ApoE and is another mechanism by which IGFBPL1 provides antiinflammatory activity. DCC is highly expressed in the brain with highest expression in neurons, and is highly expressed in neurons in the eye. DCC is a receptor of netrin, and mutations of DCC have been associated with depression, psychiatric disorders, and Parkinson’s Disease. It has been previously reported that when netrin is applied exogenously neurite outgrowth is observed. Accordingly, without wishing to be bound by theory, IGFBPLl’s ability to promote neuronal survival is believed to be based on binding to DCC, similar to its endogenous ligand netrin.

[0092] In some aspects, the disclosure provides a means for binding to DCC. In some embodiments, the means for binding to DCC is an IGFBPL1 polypeptide or IGFBPL1 fragment described herein. In some embodiments, the means for binding to DCC is a fusion protein described herein.

[0093] In some aspects, the disclosure provides a means for binding to SORT1. In some embodiments, the means for binding to SORT1 is an IGFBPL1 polypeptide or IGFBPL1 fragment described herein. In some embodiments, the means for binding to SORT1 is a fusion protein described herein.

[0094] In some aspects, the disclosure provides a means for binding to DCC and SORT1. In some embodiments, the means for binding to DCC and SORT1 is an IGFBPL1 polypeptide or IGFBPL1 fragment described herein. In some embodiments, the means for binding to DCC and SORT1 is a fusion protein described herein.

[0095] Provided herein are IGFBPL1 fragments comprising one or more domains of the wildtype IGFBPL1 protein (e.g., human wildtype IGFPBL1 protein) and fusion proteins comprising one or more domains of the wildtype IGFBPL1 protein (e.g., human wildtype IGFPBL1 protein) and a half-life extending domain. Also provided are methods of inhibiting SORT1 and / or DCC, along with methods for preferential targeting of neurons and microglia. Definitions

[0096] Terms used in the claims and specification are defined as set forth below unless otherwise specified.

[0097] As used herein, "about" will be understood by persons of ordinary skill and will vary to some extent depending on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill given the context in which it is used, "about" will mean up to plus or minus 10% of the particular value.

[0098] "Amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, g-carboxy glutamate, and O- phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that function in a manner similar to a naturally occurring amino acid.

[0099] Amino acids can be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, can be referred to by their commonly accepted single-letter codes.

[0100] An "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a predetermined amino acid sequence (an amino acid sequence of a starting polypeptide) with a second, different "replacement" amino acid residue. An "amino acid insertion" refers to the incorporation of at least one additional amino acid into a predetermined amino acid sequence. While the insertion will usually consist of the insertion of one or two amino acid residues, larger "peptide insertions,” can also be made, e.g. insertion of about three to about five or even up to about ten, fifteen, or twenty amino acid residues. The inserted residue(s) may be naturally occurring or non- naturally occurring as disclosed above. An "amino acid deletion" refers to the removal of at least one amino acid residue from a predetermined amino acid sequence.

[0101] "Polypeptide," "peptide", and "protein” are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.

[0102] "Nucleic acid" refers to deoxyribonucleotides or ribonucleoti des and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences and as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et ah, 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 conservative. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene. Polynucleotides used herein can be composed of any polyribonucleotide or polydeoxribonucleotide, which can be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can be composed of single- and double- stranded DNA, DNA that is a mixture of single- and double- stranded regions, single- and double- stranded RNA, and RNA that is mixture of single- and double- stranded regions, hybrid molecules comprising DNA and RNA that can be single- stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, the polynucleotide can be composed of triple- stranded regions comprising RNA or DNA or both RNA and DNA. A polynucleotide can also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, "polynucleotide" embraces chemically, enzymatically, or metabolically modified forms. Polynucleotides may be synthetic or isolated nucleic acid polymers including a plurality of nucleotide subunits.

[0103] As used herein, the terms “linked,” “conjugated,” “coupled,” “fused,” or “fusion,” are used interchangeably when referring to the joining together of two more elements or components or domains, by whatever means including chemical conjugation or recombinant means. Methods of chemical conjugation (e.g., using heterobifunctional crosslinking agents) are known in the art.

[0104] A peptide, polypeptide, or amino acid sequence "derived from" a designated polypeptide or protein refers to the origin of the polypeptide. Preferably, the peptide, polypeptide, or amino acid sequence which is derived from a particular sequence has an amino acid sequence that is essentially 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 which is otherwise identifiable to one of ordinary skill in the art as having its origin in the sequence. Polypeptides derived from another peptide may have one or more mutations relative to the starting polypeptide, e.g., one or more amino acid residues which have been substituted with another amino acid residue or which has one or more amino acid residue insertions or deletions.

[0105] A peptide or polypeptide can comprise an amino acid sequence which is not naturally occurring. Such variants necessarily have less than 100% sequence identity or similarity with the starting molecule. In certain embodiments, the variant will have an amino acid sequence from about 75% to less than 100% amino acid sequence identity or similarity with the amino acid sequence of the starting polypeptide, more preferably from about 80% to less than 100%, more preferably from about 85% to less than 100%, more preferably from about 90% to less than 100% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) and most preferably from about 95% to less than 100%, e.g., over the length of the variant molecule.

[0106] In certain embodiments, there is one amino acid difference between a starting polypeptide sequence and the sequence derived therefrom. Identity or similarity with respect to this sequence is defined herein as the percentage of amino acid residues in the candidate sequence that are identical (i.e., same residue) with the starting amino acid residues, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. In certain embodiments, the peptides or polypeptides are encoded by a nucleotide sequence. Nucleotide sequences of the invention can be useful for a number of applications, including: cloning, gene therapy, protein expression and purification, mutation introduction, DNA vaccination of a host in need thereof, antibody generation for, e.g., passive immunization, PCR, primer and probe generation, and the like.

[0107] As used herein, the term "operably linked" or "operably coupled" refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner.

[0108] The term "in vivo" refers to processes that occur in a living organism.

[0109] The term "mammal" or "subject" or "patient" as used herein includes both humans and non-humans and includes, but is not limited to, humans, non-human primates, canines, felines, murines, bovines, equines, and porcines. 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 the disclosure. The subject may be a vertebrate, for example, a mammal. The subject may be a human. The subjects may be symptomatic or asymptomatic. The term does not denote a particular age or sex. Adult and newborn subjects, whether male or female, are intended to be covered. A subject may include a control subject or a test subject.

[0110] The term "percent identity," in the context of two or more nucleic acid or polypeptide sequences, refer to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to persons of skill) or by visual inspection. Depending on the application, the "percent identity" can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared. For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity' for the test sequence(s) relative to the reference sequence, based on the designated program parameters.

[0111] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sei. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, PASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et ah, infra).

[0112] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et ah, J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information website.

[0113] As used herein, the term "gly-ser linker” refers to a peptide that consists of glycine and serine residues. An exemplary' gly-ser polypeptide linker comprises the amino acid sequence Ser(Gly4Ser)n. In certain embodiments, n=L In certain embodiments, n=2. In certain embodiments, n=3, i.e., Ser(Gly4Ser)3. In certain embodiments, n=4, i.e., Ser(Gly4Ser)4. In certain embodiments, n=5. In certain embodiments, n=6. In certain embodiments, n=7. In certain embodiments, n==8. In certain embodiments, n=::9. In certain embodiments, h= 10. Another exemplary gly-ser polypeptide linker comprises the amino acid sequence (Gly4Ser)n. In certain embodiments, n=l. In certain embodiments, rr::2 In certain embodiments, n=3. In certain embodiments, n=4. In certain embodiments, n=5. In certain embodiments, n=6. Another exemplary gly-ser polypeptide linker comprises the amino acid sequence (GlysSer)n. certain embodiments, n=l. In certain embodiments, n=2. In certain embodiments, n=3. In certain embodiments, n=4. In certain embodiments, n:=:5. In certain embodiments, n=6.

[0114] As used herein, the term "Fc region", ‘Fc chain” and “Fc domain” are referred to interchangeably and each refer to the portion of a native immunoglobulin formed by the respective Fc domains (or Fc moieties) of its two heavy chains. As used herein, the term "Fc domain” refers to a portion of a single immunoglobulin (Ig) heavy chain wherein the Fc domain does not comprise an Fv domain. As such, an Fc domain can also be referred to as "Ig" or "IgG." In some embodiments, an Fc domain begins in the hinge region just upstream of the papain cleavage site and ends at the C -terminus of the antibody. Accordingly, a complete Fc domain comprises at least a hinge domain, a CH2 domain, and a CHS domain. In some embodiments, an Fc domain comprises at least one of: a hinge (e.g., upper, middle, and / or lower hinge region) domain, a CH2 domain, a CHS domain, a CH4 domain, or a variant, portion, or fragment thereof. In some embodiments, an Fc domain comprises a complete Fc domain (i.e., a hinge domain, a CH2 domain, and a CHS domain). In some embodiments, an Fc domain comprises a hinge domain (or portion thereof) fused to a CH3 domain (or portion thereof). In some embodiments, an Fc domain comprises a CH2 domain (or portion thereof) fused to a CH3 domain (or portion thereof). In some embodiments, an Fc domain consists of a CH3 domain or portion thereof. In some embodiments, an Fc domain consists of a hinge domain (or portion thereof) and a CH3 domain (or portion thereof). In some embodiments, an Fc domain consists of a CH2 domain (or portion thereof) and a CH3 domain. In some embodiments, an Fc domain consists of a hinge domain (or portion thereof) and a CH2 domain (or portion thereof). In some embodiments, an Fc domain lacks at least a portion of a CH2 domain (e.g., all or part of a CH2 domain). .An Fc domain herein generally refers to a polypeptide comprising all or part of the Fc domain of an immunoglobulin heavy-chain. This includes, but is not limited to, polypeptides comprising the entire CHI, hinge, CH2, and / or CH3 domains as well as fragments of such peptides comprising only, e.g., the hinge, CH2, and CH3 domain. In some embodiments, the Fc domain is derived from an immunoglobulin of any species and / or any subtype, including, but not limited to, a human IgGl, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibody. A human IgGl constant region can be found at Uniprot P01857 and in Table 12 (i.e., SEQ ID NO: 172). The Fc domain of human IgGl can be found in Table 12 (i.e., SEQ ID NO: 173). The Fc domain encompasses native Fc and Fc variant molecules. As with Fc variants and native Fc's, the term Fc domain includes molecules in monomeric or multimeric form, whether digested from whole antibody or produced by other means. The assignment of amino acid residue numbers to an Fc domain is in accordance with the definitions of Kabat. See, e.g., 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 of Health and Human Sendees, NIH, 5th edition, Bethesda, MD vol. l:xiii-xcvi (1991); Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987); Chothia et al., Nature 342:878-883 (1989), each of which is herein incorporated by reference for all purposes. As set forth herein, it will be understood by one of ordinary skill in the art that any Fc domain may be modified such that it varies in amino acid sequence from the native Fc domain of a naturally occurring immunoglobulin molecule. In some embodiments, the Fc domain has reduced effector function (e.g., FcyR binding).

[0115] In some embodiments, the Fc domains are derived from different immunoglobulin molecules. For example, an Fc domain may comprise a CH2 and / or CH3 domain derived from an IgGl molecule and a hinge region derived from an IgG3 molecule. In another example, an Fc domain can comprise a chimeric hinge region derived, in part, from an IgGl molecule and, in part, from an IgG3 molecule. In another example, an Fc domain can comprise a chimeric hinge derived, in part, from an IgGl molecule and, in part, from an IgG4 molecule.

[0116] As used herein, the term "fusion protein" refers to a recombinant protein prepared by fusion of a multispecific variable region described herein, and a polymer (e.g., serum albumin).

[0117] The term “identity” as used herein in the context of sequences refers to a relationship between the sequences of two or more sequences, as determined by comparing the sequences. The term "identity" also means the degree of sequence relatedness between the polypeptides, as determined by the number of matches between strings of two or more amino acid residues. The percent “identity” between the two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity equals number of identical positions / total number of positions x 100), taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters. Additionally, or alternatively, the amino acid sequences disclosed herein can further be used as a “query sequence” to perform a search against public databases to, for example, identify related sequences. For example, such searches can be performed using the BLAST program of Altschul et al. (J. Mol. Biol. 215:403-10, 1990).

[0118] As used herein, the term “conservative substitution” refers to a 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 in that they have a basic sidechain, and aspartic acid and glutamic acid have similar properties in that they have an acidic side chain. In addition, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine and tryptophan have similar properties in that they have an uncharged polar side-chain, and alanine, valine, leucine, threonine, isoleucine, proline, phenylalanine and methionine have similar properties in that they have a non-polar side-chain. Also, tyrosine, phenylalanine, tryptophan and histidine have similar properties in that they have an aromatic side-chain. Thus, it will be obvious to those skilled in the art that, even when substitution of amino acid residues in groups showing similar properties as described above occurs, it will likely show no significant change in the properties of the antibody or antigen-binding fragment.

[0119] As used herein, "half-life" refers to the time taken for the serum or plasma concentration of a polypeptide to reduce by 50%, in vivo, for example due to degradation and / or clearance or sequestration by natural mechanisms. The extended-PK IGFBPL1 suitable for use in the methods disclosed herein is stabilized in vivo and its half-life increased by, e.g., fusion to an Fc region, fusion to serum albumin (e.g., HSA or MSA), through PEGylation, or by binding to serum albumin molecules (e.g., human serum albumin) which resist degradation and / or clearance or sequestration. The half-life can be determined in any manner known per se, such as by pharmacokinetic analysis. Suitable techniques will be clear to the person skilled in the art, and may for example generally involve the steps of suitably administering a suitable dose of the amino acid sequence or compound to a subject; collecting blood samples or other samples from said subject at regular intervals; determining the level or concentration of the amino acid sequence or compound in said blood sample; and calculating, from (a plot of) the data thus obtained, the time until the level or concentration of the amino acid sequence or compound has been reduced by 50% compared to the initial level upon dosing. Further details are provided in, e.g., standard handbooks, such as Kenneth, A. et al., Chemical Stability of Pharmaceuticals: A Handbook for Pharmacists and in Peters et al., Pharmacokinetic Analysis: A Practical Approach (1996). Reference is also made to Gibaldi, M. et al., Pharmacokinetics, 2nd Rev. Edition, Marcel

[0120] Dekker (1982)

[0121] IGFBPL1 Polypeptides and Fusion Proteins

[0122] In some embodiments, the disclosure provides a fusion protein comprising a human IGFBPL1 (hIGFBPLI) polypeptide or a fragment thereof, and a half-life extending domain. The hIGFBPLI polypeptide or the fragment thereof is operably linked to the half-life extending domain with or without a linker.

[0123] In some embodiments, the disclosure provides an IGFBPL1 fragment of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment is a fragment of hIGFBPLI. In some embodiments, the IGFBPL1 fragment is a fragment of murine IGFBPL1 (mIGFBPLI). In some embodiments, the IGFBPL1 fragment comprises a fragment of mIGFBPLI and a fragment of hIGFBPLI.

[0124] In some embodiments, the disclosure provides an IGFBPL1 polypeptide comprising hIGFBPLI and a fragment of mIGFBPLI. In some embodiments, the disclosure provides an IGFBPL1 polypeptide comprising mIGFBPLI and a fragment of hIGFBPLI.

[0125] IGFBPL1 Polypeptides and Fragments

[0126] Insulin like growth factor binding protein like 1 (IGFBPL1) is a soluble protein having multiple domains. Specifically, wild-type IGFBPL1 includes a thumb domain, an N-terminal domain, a Kazal domain, an Ig-like domain, and a C-terminal domain. In some embodiments, IGFBPL1 polypeptides suitable for use in the fusion proteins described herein comprise any one of the domains present in wild-type IGFBPL1. In some embodiments, IGFBPL1 polypeptides suitable for use in the fusion proteins described herein comprise any combination of the domains present in wild-type IGFBPL1.

[0127] One of ordinary skill in the art will be able to identify the domains of the IGFBPL1 polypeptide based on the IGFBPL1 polypeptide selected.

[0128] In some embodiments, the IGFPL1 polypeptides and fragments described herein are means for binding DCC. In some embodiments, the IGFPL1 polypeptides and fragments described herein are means for binding SORT1. In some embodiments, the IGFPL1 polypeptides and fragments described herein are means for binding DCC and S0RT1. Methods for determining binding of molecules are known to those of skill in the art.

[0129] In some embodiments, the thumb domain comprises the amino acid sequence set forth in SEQ ID NO: 25. In some embodiments, the thumb domain comprises an amino acid sequence 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 SEQ ID NO: 25. In some embodiments, the thumb domain comprises an amino acid sequence having 8-11 amino acid residues of 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. In some embodiments, the thumb domain comprises at least one amino acid substitution, deletion, and / or insertion 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.

[0130] In some embodiments, the N-terminal domain comprises the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the N-terminal domain comprises an amino acid sequence 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 SEQ ID NO: 2. In some embodiments, the N-terminal domain comprises an amino acid sequence having 60-70 amino acid residues of SEQ ID NO: 2. In some embodiments, the N-terminal comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the N-terminal comprises up to five amino acid substitutions, deletions, and / or insertions. In some embodiments, the N-terminal comprises at least one amino acid substitution, deletion, and / or insertion relative to SEQ ID NO: 2. In some embodiments, the N-terminal comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 2.

[0131] In some embodiments, the Kazal domain comprises the amino acid sequence set forth in SEQ ID NO: 27. In some embodiments, the Kazal domain comprises an amino acid sequence 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 SEQ ID NO: 27. In some embodiments, the Kazal domain comprises an amino acid sequence having 32-42 amino acid residues of 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. In some embodiments, the Kazal domain comprises at least one amino acid substitution, deletion, and / or insertion 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.

[0132] In some embodiments, the Ig-like domain comprises the amino acid sequence set forth in SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises an amino acid sequence 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 SEQ ID NO: 26. In some embodiments, the Ig-like domain comprises an amino acid sequence having 98-108 amino acid residues of 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. In some embodiments, the Ig-like domain comprises at least one amino acid substitution, deletion, and / or insertion 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.

[0133] In some embodiments, the C-terminal domain comprises the amino acid sequence set forth in SEQ ID NO: 3. In some embodiments, the C-terminal domain comprises an amino acid sequence 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 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-terminal comprises at least one amino acid substitution, deletion, and / or insertion. In some embodiments, the C-terminal comprises up to five amino acid substitutions, deletions, and / or insertions. In some embodiments, the C-terminal comprises at least one amino acid substitution, deletion, and / or insertion relative to SEQ ID NO: 3. In some embodiments, the C-terminal comprises up to five amino acid substitutions, deletions, and / or insertions relative to SEQ ID NO: 3.

[0134] In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the thumb domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide.

[0135] In some embodiments, the IGFBPL1 fragment comprises one to four IGFBPL1 domains selected from the N-terminal domain, the C-terminal domain, the thumb domain, the Kazal domain, and the Ig-like domain, of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises two IGFBPL1 domains selected from the N-terminal domain, the C-terminal domain, the thumb domain, the Kazal domain, and the Ig-like domain, of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises three IGFBPL1 domains selected from the N-terminal domain, the C-terminal domain, the thumb domain, the Kazal domain, and the Ig-like domain, of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises four IGFBPL1 domains selected from the N- terminal domain, the C-terminal domain, the thumb domain, the Kazal domain, and the Ig-like domain, of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises no more than four of the N-terminal domain, the C-terminal domain, the thumb domain, the Kazal domain, and the Ig-like domain, of an IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises more than one copy of an IGFBPL1 domain described herein.

[0136] In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain and the thumb domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain and the C-terminal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain and the thumb domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C- terminal domain and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the thumb domain and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the thumb domain and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the Kazal domain and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide.

[0137] In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the Kazal domain, and the thumb domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C-terminal domain, and the thumb domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C-terminal domain, and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C-terminal domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the thumb domain, and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the Ig-like domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain, the thumb domain, and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain, the thumb domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the Kazal domain, the thumb domain, and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain, the Ig-like domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide.

[0138] In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C- terminal domain, the thumb domain, and the Ig-like domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C-terminal domain, the thumb domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the C-terminal domain, the Ig-like domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the N-terminal domain, the thumb domain, the Ig-like domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide. In some embodiments, the IGFBPL1 fragment comprises the C-terminal domain, the thumb domain, the Ig-like domain, and the Kazal domain of an IGFBPL1 polypeptide, provided the IGFBPL1 fragment does not comprise other domains of the IGFBPL1 polypeptide.

[0139] In some embodiments, the IGFBPL1 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 to 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 IGFBPL1 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 to an amino acid sequence selected from SEQ ID Nos: 2-3, 25-27, 37-50, 158, and 160-179. In some embodiments, the IGFBPL1 fragment comprises 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 IGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID Nos: 2-3, 25-27, 37-50, 158, and 160-179. In some embodiments, the IGFBPL1 fragment consists of 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 IGFBPL1 fragment consists of an amino acid sequence selected from SEQ ID Nos: 2-3, 25-27, 37-50, 158, and 160-179.

[0140] In some embodiments, an IGFBPL1 polypeptide provided herein is a chimeric IGFBPL1 polypeptide comprising at least one IGFBPL1 domain of a human IGFBPL1 polypeptide and at least one IGFBPL1 domain of a non-human IGFBPL1 polypeptide. In some embodiments, the non-human IGFBPL1 polypeptide is a murine IGFBPL1 polypeptide. In some embodiments, a chimeric IGFBPL1 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 to an amino acid sequence selected from SEQ ID Nos: 151-157. In some embodiments, a chimeric IGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID Nos: 151-157. In some embodiments, a chimeric IGFBPL1 polypeptide consists of an amino acid sequence selected from SEQ ID Nos: 151-157.

[0141] In some embodiments, a fusion protein described herein comprises an IGFBPL1 polypeptide or IGFBPL1 fragment described herein.

[0142] In some embodiments, the fusion protein comprises the N-terminal domain of an IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the N-terminal domain of an IGFBPL1 polypeptide and does not comprise the C-terminal domain of the IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C-terminal domain of an IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C- terminal domain of an IGFBPL1 polypeptide and does not comprise the N-terminal domain of the IGFBPL1 polypeptide.

[0143] In some embodiments, the fusion protein comprises the N-terminal domain of a human IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the N-terminal domain of a human IGFBPL1 polypeptide and does not comprise the C-terminal domain of the human IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C- terminal domain of a human IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C-terminal domain of a human IGFBPL1 polypeptide and does not comprise the N-terminal domain of the human IGFBPL1 polypeptide.

[0144] In some embodiments, the fusion protein comprises the N-terminal domain of a human IGFBPL1 wild-type polypeptide. In some embodiments, the fusion protein comprises the N- terminal domain of a human wild-type IGFBPL1 polypeptide and does not comprise the C- terminal domain of the human wild-type IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C-terminal domain of a human wild-type IGFBPL1 polypeptide. In some embodiments, the fusion protein comprises the C-terminal domain of a human wild-type IGFBPL1 polypeptide and does not comprise the N-terminal domain of the human wild-type IGFBPL1 polypeptide.

[0145] In some embodiments, the fusion protein comprises the N-terminal domain of a human IGFBPL1 polypeptide as set forth in SEQ ID NO: 2. In some embodiments, the fusion protein comprises the N-terminal domain of a human IGFBPL1 polypeptide as set forth in SEQ ID NO: 2 and does not comprise the C-terminal domain of the human IGFBPL1 polypeptide as set forth in SEQ ID NO: 3. In some embodiments, the fusion protein comprises the C-terminal domain of a human IGFBPL1 polypeptide as set forth in SEQ ID NO: 3. In some embodiments, the fusion protein comprises the C-terminal domain of a human IGFBPL1 polypeptide as set forth in SEQ ID NO: 3 and does not comprise the N-terminal domain of the human IGFBPL1 polypeptide as set forth in SEQ ID NO: 2.

[0146] In some embodiments, the fusion protein comprises a domain of an IGFBPL1 polypeptide that binds to SORT1. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to SORT1 is an Ig-like domain. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to SORT1 is an Ig-like domain and a Kazal domain. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to SORT1 is an Ig-like domain, a Kazal domain, and a C-terminal domain. In some embodiments, the fusion protein comprises a domain of an IGFBPL1 polypeptide that binds to SORT1 and does not bind to DCC.

[0147] In some embodiments, the fusion protein comprises a domain of an IGFBPL1 polypeptide that binds to DCC. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to DCC is an N-terminal domain. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to DCC is an N-terminal domain and a thumb domain. In some embodiments, the domain of an IGFBPL1 polypeptide that binds to DCC is an N-terminal domain, a thumb domain, and a Kazal domain. In some embodiments, the fusion protein comprises a domain of an IGFBPL1 polypeptide that binds to DCC and does not bind to SORT1.

[0148] In some embodiments, the fusion protein comprises domains of an IGFBPL1 polypeptide that binds DCC and SORT1.

[0149] In some embodiments, the fusion protein comprises a fragment of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises one or more domain of an IGFBPL1 polypeptide. In some embodiments, the fragment does not comprise all domains of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises a portion of one or more domains of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the thumb domain of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the thumb domain of a human IGFBPL1 polypeptide. In some embodiments, the fragment comprises a thumb domain comprising the amino acid sequence of SEQ ID NO: 25. In some embodiments, the fragment comprises the N-terminal domain of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the N-terminal domain of a human IGFBPL1 polypeptide. In some embodiments, the fragment comprises the N-terminal domain comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the fragment comprises the Kazal domain of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the Kazal domain of a human IGFBPL1 polypeptide. In some embodiments, the fragment comprises the Kazal domain comprising the amino acid sequence of SEQ ID NO: 27. In some embodiments, the fragment comprises the Ig-like domain of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the Ig-like domain of a human IGFBPL1 polypeptide. In some embodiments, the fragment comprises the Ig-like domain comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the fragment comprises the C-terminal domain of an IGFBPL1 polypeptide. In some embodiments, the fragment comprises the C-terminal domain of a human IGFBPL1 polypeptide. In some embodiments, the fragment comprises the C-terminal domain comprising the amino acid sequence of SEQ ID NO: 3.

[0150] In some embodiments, the fusion protein comprises a fragment of an hIGFBPLI polypeptide. In some embodiments, the fragment comprises amino acid residues P36-M278 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues P36-A109 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-E152 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues P36-E152 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues V107-E152 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues C108-E152 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues V107-M278 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues C108-M278 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues V107-S264 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues C108-S264 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues E152-M278 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues E152-S264 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-Q174 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-G104 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-G112 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-G119 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues L26-A95 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues K37-G104 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues K37-G112 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues K37-G119 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues K37-A95 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues C151A-T259 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues C151A-R277 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues A154-T259 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues A154-R277 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues G71-T259 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues G71-R277 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues A96-T259 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues K37-E152 of an IGFBPL1 polypeptide by straight numbering. In some embodiments, the fragment comprises amino acid residues E152-R277 of an IGFBPL1 polypeptide by straight numbering.

[0151] 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.

[0152] In some embodiments, the fragment comprises at least two domains of IGFBPL1 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, a fragment comprising at least two domains of IGFBPL1 operably linked via a linker comprises an amino acid sequence selected from SEQ ID Nos: 175- 177.

[0153] In some embodiments, the fragment comprises mutation C108S. In some embodiments, the fragment comprises mutation V107X. In some embodiments, the fragment comprises mutation V107T. In some embodiments, the fragment comprises mutation C108S. In some embodiments, the fragment comprises the mutation C151A.

[0154] In some embodiments, the fragment comprises an amino acid sequence selected from SEQ ID NOs: 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 to any one of the amino acid sequences set forth in SEQ ID Nos: 37-50.

[0155] In some embodiments, the fragment comprises an amino acid sequence selected from SEQ ID NOs: 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 to any one of the amino acid sequences set forth in SEQ ID Nos: 158 and 160-179.

[0156] In some embodiments, the fragment comprises an amino acid sequence selected from

[0157] SEQ ID NOs: 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 to any one of the amino acid sequences set forth in SEQ ID Nos: 37-50, 158 and 160-179.

[0158] In some embodiments, the fusion protein comprises a hIGFBPLI polypeptide comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the fusion protein comprises a hIGFBPLI polypeptide comprising a sequence that is 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 set forth in SEQ ID NO: 10. In some embodiments, the fusion protein a hIGFBPLI polypeptide comprising the sequence set forth in SEQ ID NO: 10 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0159] In some embodiments, the fusion protein comprises a hIGFBPLI polypeptide comprising the amino acid sequence of SEQ ID NO: 11. In some embodiments, the fusion protein comprises a hIGFBPLI polypeptide comprising a sequence that is 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 set forth in SEQ ID NO: 11. In some embodiments, the fusion protein comprises a hIGFBPLI polypeptide comprising the sequence set forth in SEQ ID NO: 11 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0160] In some embodiments, the IGFBPL1 polypeptide or the fragment thereof comprises a mutation that alters glycosylation. In some embodiments, the glycosylation mutation is at position N166 of an hIGFBPLI polypeptide. In some embodiments, the glycosylation mutation is N166D. In some embodiments, an IGFBPL1 polypeptide having an N166D mutation is set forth in SEQ ID NO: 87. In some embodiments, the glycosylation mutation is N166Q. In some embodiments, an IGFBPL1 polypeptide having an N166Q mutation is set forth in SEQ ID NO: 88. In some embodiments, the glycosylation mutation is at position S268 of an hIGFBPLI. In some embodiments, the glycosylation mutation is S268A. In some embodiments, an IGFBPL1 polypeptide having an S268A mutation is set forth in SEQ ID NO: 89. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof comprises glycosylation mutations at positions N166 and S268. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof comprises glycosylation mutations N166D and S268A. In some embodiments, an IGFBPL1 polypeptide having an N166D and S268A mutation is set forth in SEQ ID NO: 90. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof comprises glycosylation mutations N166Q and S268A. In some embodiments, an IGFBPL1 polypeptide having an N166Q and S268A mutation is set forth in SEQ ID NO: 91.

[0161] In some embodiments, the IGFBPL1 fragments described herein do not comprise a halflife extending domain and are useful in compositions and methods disclosed herein.

[0162] Half-Life Extending Domains

[0163] In some embodiments, the fusion proteins disclosed herein comprise a half-life extending domain. In some embodiments, a half-life extending domain increases circulation half-life of the IGFBPL1 polypeptide or the fragment thereof.

[0164] Half-life extending domains are well known in the art, see, e.g., Strohl, BioDrugs. 2015; 29(4): 215-239 and Zaman et al., Journal of Controlled Release 301 (2019) 176-189, each of which is incorporated herein in its entirety for examples of half-life extending domains that may be used in the fusion proteins provided herein. Half-life extending domains are also described in WO 2016 / 025647 (incorporated herein by this reference) as provided below.

[0165] A half-life extending domain that may be used in the fusion proteins described herein includes any protein domain that may be fused to an IGFBPL1 domain to extend its half-life. Examples of half-life extending domains include Fc domains, albumin domains, transferrin domains, C-terminal peptides (e.g., of chorionic gonadotropin P-chain), elastin-like peptides, non-exact repeat peptide sequences, proline-alanine-serine polymers, repeat sequences of glycine rich polypeptides, and gelatin-like proteins.

[0166] Fc Domains

[0167] In some embodiments, the half-life extending moiety is the Fc domain of an immunoglobulin. A variety of Fc domain gene sequences (e.g., mouse and human constant region gene sequences) are available in the form of publicly accessible deposits. Constant region domains comprising an Fc domain sequence can be selected lacking a particular effector function and / or with a particular modification to reduce immunogenicity. Many sequences of antibodies and antibody-encoding genes have been published and suitable Fc domain sequences (e.g. hinge, CH2, and / or CH3 sequences, or portions thereof) can be derived from these sequences using art recognized techniques. The genetic material obtained using any of the foregoing methods may then be altered or synthesized to obtain polypeptides suitable for use in the methods disclosed herein. It will further be appreciated that the scope of this invention encompasses alleles, variants and mutations of constant region DNA sequences.

[0168] The half-life extending domain may be the Fc domain of any type of immunoglobulin, including IgA, IgD, IgE, IgG, and IgM. The half-life extending moiety may be the Fc domain of any subtype of immunoglobulin, including IgGl, IgG2, IgG3, or IgG4. In some embodiments, the half-life extending domain is the Fc domain of human IgGl.

[0169] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO:

[0170] 4. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 4. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 4 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0171] In some embodiments the Fc domain comprises the amino acid sequence of SEQ ID NO:

[0172] 5. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 5. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 5 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0173] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO:

[0174] 6. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 6. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 6 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0175] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO:

[0176] 7. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 7. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 7 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0177] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO:

[0178] 8. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 8. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 8 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 9. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 9. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 9 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0179] In some embodiments, the Fc domain comprises the amino acid sequence of SEQ ID NO: 137. In some embodiments, the Fc domain comprises a sequence that is 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 set forth in SEQ ID NO: 137. In some embodiments, the Fc domain comprises the sequence set forth in SEQ ID NO: 137 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0180] In some embodiments, an Fc domain employed in the fusion protein disclosed herein is altered or modified, e.g., by amino acid mutation (e.g., addition, deletion, or substitution). As used herein, the term "Fc domain variant" refers to an Fc domain having at least one amino acid modification, such as an amino acid substitution, as compared to the wild-type Fc from which the Fc domain is derived. For example, wherein the Fc domain is derived from a human IgGl antibody, a variant comprises at least one amino acid mutation (e.g., substitution) as compared to a wild type amino acid at the corresponding position of the human IgGl Fc region.

[0181] In some embodiments, the Fc variant comprises a substitution at an amino acid position located in a hinge domain or portion thereof. In certain embodiments, the Fc variant comprises a substitution at an amino acid position located in a CH2 domain or portion thereof. In certain embodiments, the Fc variant comprises a substitution at an amino acid position located in a CH3 domain or portion thereof. In certain embodiments, the Fc variant comprises a substitution at an amino acid position located in a CH4 domain or portion thereof.

[0182] In some embodiments, the fusion protein comprises an Fc variant comprising more than one amino acid substitution. The fusion protein may comprise, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid substitutions. Preferably, the amino acid substitutions are spatially positioned from each other by an interval of at least 1 amino acid position or more, for example, 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 positioned apart from each other by an interval of at least 5, 10, 15, 20, or 25 amino acid positions or more. In some embodiments, an Fc domain includes changes in the region between amino acids 234- 238, including the sequence LLGGP at the beginning of the CH2 domain. In some embodiments, an Fc variant alters Fc mediated effector function, particularly ADCC, and / or decrease binding avidity for Fc receptors. In some embodiments, sequence changes closer to the CH2-CH3 junction, at positions such as K322 or P331 can eliminate complement mediated cytotoxicity and / or alter avidity for FcR binding. In some embodiments, an Fc domain incorporates changes at residues P238 and P331, e.g., changing the wild type prolines at these positions to serine. In some embodiments, alterations in the hinge regi on at one or more of the three hinge cysteines, to encode CCC, SCC, SSC, SCS, or SSS at these residues can also affect FcR binding and molecular homogeneity, e.g., by elimination of unpaired cysteines that may destabilize the folded protein.

[0183] Other amino acid mutations in the Fc domain are contemplated to reduce binding to the Fc gamma receptor and Fc gamma receptor subtypes. For example, mutations at positions 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 283, 285, 298,

[0184] 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 312, 315, 322, 324, 327, 329, 330,

[0185] 331, 333, 334, 335, 337, 338, 340, 356, 360, 373, 376, 378, 379, 382, 388, 389, 398, 414, 416,

[0186] 419, 430, 434, 435, 437, 438 or 439 of the Fc region can alter binding as described in U.S. Pat.

[0187] No. 6,737,056, issued May 18, 2004, incorporated herein by reference in its entirety. This patent reported that changing Pro331 in IgG3 to Ser resulted in six-fold lower affinity as compared to unmutated IgG3, indicating the involvement of Pro331 in Fc gamma RI binding. In addition, amino acid modifications at positions 234, 235, 236, and 237, 297, 318, 320 and 322 are disclosed as potentially altering receptor binding affinity in U.S. 5,624,821, issued April 29, 1997 and incorporated herein by reference in its entirety.

[0188] In some embodiments, the Fc domain comprises an amino acid substitution atN297. In some embodiments, the amino acid substitution is N297D. In some embodiments, the amino acid substitution is N297Q. In some embodiments, an Fc domain with an N297D mutation comprises the amino acid sequence set forth in SEQ ID NO: 92. In some embodiments, an Fc domain with an N297Q mutation comprises the amino acid sequence set forth in SEQ ID NO: 93.

[0189] Further mutations contemplated for use include, e.g., those described in U.S. Pat. App. Pub. No. 2006 / 0235208, published October 19, 2006, and incorporated herein by reference in its entirety. This publication describes Fc variants that exhibit reduced binding to Fc gamma receptors, reduced antibody dependent cell-mediated cytotoxicity, or reduced complement dependent cytotoxicity, that comprise 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, 327R, 328R, 329K, 3301, 330L, 330N, 330P, 330R, and 33 IL (numbering is according to the EU index), as well as 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 contemplated for use as described in this publication 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, 328M, 328N, 328Q, 328T, 328V, 328Y, 3301, 330L, 330Y, 332D, 332E, 335D, an insertion of G between positions 235 and 236, an insertion of A between positions 235 and 236, an insertion of S between positions 235 and 236, an insertion of T between positions 235 and 236, an insertion of N between positions 235 and 236, an insertion of D between positions 235 and 236, an insertion of V between positions 235 and 236, an insertion of L between positions 235 and 236, an insertion of G between positions 235 and 236, an insertion of A between positions 235 and 236, an insertion of S between positions 235 and 236, an insertion of T between positions 235 and 236, an insertion of N between positions 235 and 236, an insertion of D between positions 235 and 236, an insertion of V between positions 235 and 236, an insertion of L between positions 235 and 236, an insertion of G between positions 297 and 298, an insertion of A between positions 297 and 298, an insertion of S between positions 297 and 298, an insertion of D between positions 297 and 298, an insertion of G between positions 326 and 327, an insertion of A between positions 326 and 327, an insertion of T between positions 326 and 327, an insertion of D between positions 326 and 327, and an insertion of E between positions 326 and 327 (numbering is according to the EU index). Additionally, mutations described in U.S. Pat. App. Pub. No. 2006 / 0235208 include 227G / 332E, 234D / 332E, 234E / 332E, 234Y / 332E, 2341 332E, 234G / 332E, 235I7332E, 235S / 332E, 235D / 332E, 235E / 332E,

[0190] 236S / 332E,236A / 332E, 236S / 332D, 236A / 332D, 239D / 268E, 246H / 332E, 255Y / 332E, 258H / 332E, 260H / 332E, 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 / 268 E / 327A, 239D / 332E / 268E / 330Y / 327A, Insert 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>326-3277332E, 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 (numbering according to the EU index) are contemplated for use. The mutant L234A / L235A is described, e.g., in U.S. Pat. App. Pub. No. 2003 / 0108548, published June 12, 2003 and incorporated herein by reference in its entirety. In some embodiments, the described modifications are included either individually or in combination. In some embodiments, the mutation is D265A in human IgGl.

[0191] In some embodiments, the Fc domain comprises the mutations L234A and L235A, with numbering relative to the Kabat numbering scheme of IgG. In some embodiments, the Fc domain comprises the mutations M252Y, S254T, and T256E, with numbering relative to the Kabat numbering scheme of IgG.

[0192] In some embodiments, the fusion protein comprises an amino acid substitution to an Fc domain which alters antigen-independent effector functions of the polypeptide, in particular the circulating half-life of the polypeptide.

[0193] In some embodiments, the fusion proteins disclosed herein comprises an Fc variant comprising an amino acid substitution which alters the antigen-dependent effector functions of the polypeptide, in particular ADCC or complement activation, e.g., as compared to a wild type Fc region. Such fusion proteins exhibit decreased binding to FcR gamma when compared to wild-type polypeptides and, therefore, mediate reduced effector function. Fc variants with decreased FcR gamma binding affinity are expected to reduce effector function, and such molecules are also useful, for example, for treatment of conditions in which target cell destraction is undesirable, e.g., where normal cells may express target molecules, or where chronic administration of the polypeptide might result in unwanted immune system activation.

[0194] In some embodiments, the fusion proteins exhibit altered binding to an activating FcyR (e.g. Fcyl, Fcylla, or FcyRIIIa). In certain embodiments, the fusion proteins exhibit altered binding affinity to an inhibitory FcyR (e.g. FcyRUb). Exemplary amino acid substitutions which altered FcR or complement binding activity are disclosed in International PCT Publication No. W005 / 063815 which is incorporated by reference herein.

[0195] The fusion proteins disclosed herein may also comprise an amino acid substitution which alters the glycosylation of the extended-PK IL -2. For example, the Fc domain of the fusion protein may comprise an Fc domain having a mutation leading to reduced glycosylation (e.g., N- or O-linked glycosylation) or may comprise an altered glycoform of the wild-type Fc domain (e.g., a low fucose or fucose-free glycan). In some embodiments, the fusion protein has an amino acid substitution near or within a glycosylation motif, for example, an N-linked glycosylation motif that contains the amino acid sequence NXT or NXS. Exemplary amino acid substituti ons which reduce or alter glycosylation are disclosed in W005 / 018572 and US2007 / 0111281, the contents of which are i ncorporated by reference herein. In some embodiments, the fusi on proteins disclosed herein comprise at least one Fc domain having engineered cysteine residue or analog thereof which is located at the solvent-exposed surface. In some embodiments, the fusi on proteins disclosed herein comprise an Fc domain comprising at least one engineered free cysteine residue or analog thereof that is substantially free of disulfide bonding with a second cysteine residue. Any of the above engineered cysteine residues or analogs thereof may subsequently be conjugated to a functional domain using art-recognized techniques (e.g., conjugated with a thiol-reactive heterobifunctional linker).

[0196] In some embodiments, the fusion proteins disclosed herein may comprise a genetically fused Fc domain having two or more of its constituent Fc domains independently selected from the Fc domains described herein. In some embodiments, the Fc domains are the same. In certain embodiments, at least two of the Fc domains are different. For example, the Fc domains of the fusion proteins disclosed herein comprise the same number of amino acid residues or they may differ in length by one or more amino acid residues (e.g., by about 5 amino acid residues (e.g., 1,

[0197] 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 proteins disclosed herein may differ in sequence 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,

[0198] 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).

[0199] PEGylation

[0200] In some embodiments, the half-life extending domain is a polyethylene glycol (PEG) domain. PEGylation is well known in the art to confer increased circulation half-life to proteins. Methods of PEGylation are well known and disclosed in, e.g., US7,610,156, US7, 847,062, all of which are hereby incorporated by reference.

[0201] PEG is a well-known, water soluble polymer that is commercially available or can be 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, pages 138- 161). The term "PEG" is used broadly to encompass any polyethylene glycol molecule, without regard to size or to modification at an end of the PEG, and can be represented by the formula: X — 0(CH2CH20)n-iCH2CH20H, where n is 20 to 2300 and X is H or a terminal modification, e.g., a Ci-4 alkyl. In certain embodiments, the PEG suitable for use in the methods disclosed herein terminates on one end with hydroxy or methoxy, i.e., X is H or CEb ("methoxy PEG"). PEG can contain further chemical groups which are necessary for binding reactions; which results from the chemical synthesis of the mol ecule, or which is a spacer for optimal distance of parts of the molecule. In addition, such a PEG can consist of one or more PEG side-chains which are linked together. PEGs with more than one PEG chain are called multiarmed or branched PEGs. Branched PEGs can be prepared, for example, by the addition of polyethylene oxide to various polyols, including glycerol, pentaerythriol, and sorbitol. For example, a four-armed branched PEG can be prepared from pentaerythriol and ethylene oxide. Branched PEG are described in, for example, EP- A 0 473 084 and USS, 932,462, both of which are hereby incorporated by reference. One form of PEGs includes two PEG side-chains (PEG2) linked via the primary amino groups of a lysine (Monfardini et al.. Bioconjugate Chem 1995;6:62-9). In some embodiments, pegylated IGFBPL1 is produced by site-directed pegylation, particularly by conjugation of PEG to a cysteine moiety at the N- or C-terminus. A PEG moiety may also be attached by other chemistry, including by conjugation to amines. PEG conjugation to peptides or proteins generally involves the activation of PEG and coupling of the activated PEG-intermediates directly to target proteins / peptides or to a linker, which is subsequently activated and coupled to target proteins / peptides (see Abuchowski et al., JBC 1977;252:3571 and JBC 1977;252:3582, and Harris et. al., in: Polyethylene glycol) Chemistry: Biotechnical and Biomedical Applications; (J. M. Harris ed.) Plenum Press: New York, 1992; Chap. 21 and 22). A variety of molecular mass forms of PEG can be selected, e.g., from about 1,000 Daltons (Da) to 100,000 Da (n is 20 to 2300), for conjugating to IGFBPL1 . The number of repeating units ”n" in the PEG is approximated for the molecular mass described in Daltons. It is preferred that the combined molecular mass of PEG on an activated linker is suitable for pharmaceutical use. Thus, in one embodiment, the molecular mass of the PEG molecules does not exceed 100,000 Da. For example, if three PEG molecules are attached to a linker, where each PEG molecule has the same molecular mass of 12,000 Da (each n is about 270), then the total molecular mass of PEG on the linker is about 36,000 Da (total n is about 820). The molecular masses of the PEG attached to the linker can also be different, e.g., of three molecules on a linker two PEG molecules can be 5,000 Da each (each n is about 110) and one PEG molecule can be 12,000 Da (n is about 270).

[0202] One skilled in the art can select a suitable molecular mass for PEG, e.g., based on how the pegylated IGFBPL1 will be used therapeutically, the desired dosage, circulation time, resistance to proteolysis, immunogenicity, and other considerations. For a discussion of PEG and its use to enhance the properties of proteins, see N. V. Katre, Advanced Drug Deliver}' Reviewsl993;10:91-l 14.

[0203] In some embodiments, PEG molecules may be activated to react with amino groups on IL -2 such as with lysines (Bencham C. O. et al.. Anal. Biochem., 131, 25 (1983), Veronese, F. M. 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, 1 19-128 (1990)).

[0204] In some embodiments, carbonate esters of PEG are used to form the PEG-IL-2 conjugates. N,N'-disuccinimidylcarbonate (DSC) may be used in the reaction with PEG to form active mixed PEG-succinimidyl carbonate that may be subsequently reacted with a nucleophilic group of a linker or an amino group of IL-2 (see U.S. Pat. No. 5,281,698 and U.S. Pat. No. 5,932,462). In a similar type of reaction, 1,1'- (dibenzotri azolyl)carbonate and di-(2- pyridyl)carbonate may be reacted with PEG to form PEG-benzotriazolyl and PEG-pyridyl mixed carbonate (U.S. Pat. No. 5,382,657), respectively. Pegylation of IGFBPLI can be performed according to the methods of the state of the art, for example by reaction of IGFBPLI with electrophilically active PEGs (Shearwater Corp., USA, www.shearwatercorp.com). Preferred PEG reagents suitable for use in the methods disclosed herein are, e.g., N-hydroxysuccinimidyl propionates (PEG-SPA), butanoates (PEG-SBA), PEG- succinimidyl propionate or branched N- hydroxy succinimides such as mPEG2-NHS (Monfardini, C, et al., Bioconjugate Chetn. 6 (1995) 62-69).

[0205] In some embodiments, pegylated IGFBPLI comprise one or more PEG molecules covalently attached to a linker.

[0206] Other Half-Life Extending Domains

[0207] In some embodiments, the half-life extending domain is a serum albumin, or fragments thereof. Methods of fusing serum albumin to proteins are disclosed in, e.g., US2010 / 0144599, US2007 / 0048282, and US2011 / 0020345, which are herein incorporated by reference in their entirety. In some embodiments, the half-life extending domain is human serum albumin (HSA), or variants or fragments thereof, such as those disclosed in US 5,876,969, WO 2011 / 124718, WO 2013 / 075066, and WO 2011 / 0514789.

[0208] In some embodiments, the serum albumin comprises the amino acid sequence of SEQ ID NO: 181. In some embodiments, the serum albumin comprises a sequence that is 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 set forth in SEQ ID NO: 181. In some embodiments, the serum albumin comprises the sequence set forth in SEQ ID NO: 181 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0209] In some embodiments, the half-life extending domain is a serum albumin binding protein such as those described in US2005 / 0287153, US2007 / 0003549, US2007 / 0178082, US2007 / 0269422, US2010 / 0113339, W02009 / 083804, and W02009 / 133208, which are herein incorporated by reference in their entirety. In some embodiments, the half-life extending domain is transferrin, as disclosed in US 7,176,278 and US 8,158,579, which are herein incorporated by reference in their entirety.

[0210] In some embodiments, the half-life extending domain is a serum immunoglobulin binding protein such as those disclosed in US2007 / 0178082, which is herein incorporated by reference in its entirety.

[0211] In some embodiments, the half-life extending domain is a fibronectin (Fn)-based scaffold domain protein that binds to serum albumin, such as those disclosed in US2012 / 0094909, which is herein incorporated by reference in its entirety. Methods of making fibronectin-based scaffold domain proteins are also disclosed in US2012 / 0094909. A non-limiting example of a Fn3-based extended-PK group is Fn3(HSA), i.e., a Fn3 protein that binds to human serum albumin.

[0212] Linkers

[0213] In some embodiments, the IGFBPL1 polypeptide or the fragment thereof is operably linked to the half-life extending domain via a linker.

[0214] Linkers that may be used to connect domains of fusion proteins are well known in the art, see, e.g., Chen etal., Adv Drug Deliv Rev. 2013 Oct;65(10): 1357-69.

[0215] In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a synthetic linker. As used herein, the term "synthetic" with respect to a polypeptide linker includes peptides (or polypeptides) which comprise an amino acid sequence (which may or may not be naturally occurring) that is linked in a linear sequence of amino acids to a sequence (which may or may not be naturally occurring) (e.g., an ABP sequence) to which it is not naturally linked in nature. For example, the polypeptide linker may comprise non-naturally occurring polypeptides which are modified forms of naturally occurring polypeptides (e.g., comprising a mutation such as an addition, substitution or deletion) or which comprise a first amino acid sequence (which may or may not be naturally occurring).

[0216] In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a cleavable linker. Examples of cleavable linkers include cyclopeptide linkers, disulfide linkers, and linkers that comprise a sequence susceptible to protease cleavage. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a linker comprising a tobacco etch virus (TEV) cleavage site. An illustrative sequence of a TEV cleavable linker is GGGSENLYFQS (SEQ ID NO: 12). In some embodiments, a TEV cleavable linker comprises the amino acid sequence of SEQ ID NO: 182.

[0217] In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a flexible linker. Flexible linkers generally comprise small non-polar amino acids such as glycine, or polar amino acids such as serine and / or threonine. An example of a flexible linker is a GS linker, e.g., a G4S linker. A G4S linker comprises an amino acid sequence of the formula (Gly4Ser)n, wherein n is a positive integer (e.g., 1, 2, 3, 4, or 5). In certain embodiments the gly / ser linker is (Gly4Ser)i. In certain embodiments the gly / ser linker is (Gly4Ser)2. In certain embodiments the gly / ser linker is (Gly4Ser)3. In certain embodiments the gly / ser linker is (Gly4Ser)4. In certain embodiments the gly / ser linker is (Gly4Ser)s. In some embodiments, two or more gly-ser linker are incorporated in series in a polypeptide linker. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a G4S linker. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain are connected by a GS linker. Illustrative sequences of GS linkers 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.

[0218] Other linkers that are suitable for use in fusion proteins described herein are known in the art, for example, the serine-rich linkers disclosed in US 5,525,491, the helix forming peptide linkers (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 linkers disclosed in Chen et ah, Mol Pharm 2011;8:457-65, i.e., the dipeptide linker LE, a thrombin- sensitive disulfide cyclopeptide linker, and the alpha-helix forming linker LEA(EAAAK)4ALEA(EAAAK)4ALE (SEQ ID NO: 24).

[0219] In some embodiments, the linker is an XTEN-13k linker. In some embodiments, the XTEN-13k linker comprises the amino acid sequence of SEQ ID NO: 180.

[0220] Fusion Protein Structure & Exemplary Fusion Proteins

[0221] The IGFBPL1 polypeptide or the fragment thereof and the half-life extending domain may be arranged in any order in the fusion proteins provided herein. The means for binding DCC, SORT1, or both DCC and SORT1, and the half-life extending domain may be arranged in any order in the fusion proteins provided herein. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof is N-terminal to the half-life extending domain. In some embodiments, the IGFBPL1 polypeptide or the fragment thereof is C-terminal to the half-life extending domain.

[0222] In some embodiments, two fusion proteins provided herein dimerize. It will be apparent to a person of skill in the art that the sequences of the fusion protein domains may be manipulated to stabilize the dimerization. For example, in some embodiments, knob-in-hole design may be used to ensure dimerization of the fusion proteins. SEQ ID Nos: 5 and 8 are illustrative “knob” Fc sequences and SEQ ID Nos: 6 and 9 are illustrative “hole” Fc sequences. Methods of achieving knob-in-hole modifications are known in the art, see, e.g., Liu et al., Front. Immunol., 26 January 2017 Sec. Vaccines and Molecular Therapeutics Volume 8 - 2017.

[0223] The fusion protein dimers described herein may be monovalent or multivalent. A multivalent fusion protein dimer has more than one half-life extending moiety per each IGFBPL1 polypeptide or the fragment thereof.

[0224] 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.

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

[0226] 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.

[0227] 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. If a fusion protein dimer is a bivalent fusion protein, the half-life extending domains may be attached in trans or in cis orientation. In some embodiments, the fusion protein is a bivalent N-terminal and C-terminal trans-fusion protein. In some embodiments, the fusion protein is a bivalent N-terminal and C-terminal cis-fusion protein.

[0228] 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 Fc-fusion protein. In some embodiments, the fusion protein is a bivalent 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.

[0229] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 15. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 15 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0230] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 16. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 16 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0231] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 17. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 17 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 18. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 18 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0232] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 19. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 19 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0233] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 20. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 20 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0234] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 21. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 21 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0235] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 22. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 22 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 51. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 51 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0236] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 52. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 52 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0237] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 53. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 53 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0238] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 54. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 54 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0239] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 55. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 55 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 56. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 56 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0240] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 57. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 57 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0241] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 58. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 58 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0242] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 59. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 59 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0243] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 60. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 60 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 61. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 61 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0244] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 62. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 62 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0245] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 63. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 63 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0246] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 64. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 64 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0247] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 65. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 65 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 66. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 66 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0248] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 67. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 67 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0249] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 68. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 68 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0250] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 69. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 69 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0251] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 70. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 70 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 71. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 71 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0252] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 72. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 72 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0253] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 73. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 73 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0254] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 74. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 74 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0255] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 75. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 75 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 76. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 76 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0256] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 77. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 77 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0257] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 78. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 78 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0258] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 79. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 79 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0259] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 80. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 80 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 81. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 81 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0260] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 82. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 82 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0261] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 83. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 83 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0262] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 84. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 84 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0263] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 85. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 85 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 86. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 86 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0264] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 94. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 94 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0265] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 95. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 95 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0266] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 96. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 96 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0267] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 97. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 97 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 98. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 98 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0268] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 99. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 99 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0269] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 100. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 100 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0270] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 101. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 101 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0271] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 102. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 102 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 103. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 103 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0272] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 104. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 104 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0273] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 105. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 105 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0274] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 106. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 106 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0275] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 107. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 107 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 108. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 108 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0276] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 109. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 109 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0277] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 110. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 110 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0278] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 111. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 111 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0279] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 112. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 112 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 113. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 113 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0280] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 114. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 114 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0281] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 115. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 115 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0282] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 116. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 116 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0283] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 117. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 117 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 118. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 118 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0284] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 119. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 119 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0285] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 120. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 120 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0286] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 121. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 121 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0287] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 122. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 122 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 123. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 123 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0288] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 124. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 124 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0289] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 125. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 125 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0290] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 126. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 126 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0291] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 127. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 127 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 128. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 128 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0292] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 129. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 129 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0293] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 130. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 130 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0294] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 131. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 131 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0295] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 132. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 132 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 133. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 133 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0296] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 134. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 134 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0297] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 135. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 135 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0298] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 136. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 136 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0299] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 138. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 138 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 139. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 139 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0300] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 140. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 140 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0301] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 142. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 142 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0302] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 144. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 144 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0303] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 145. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 145 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 146. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 146 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0304] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 147. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 147 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0305] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 148. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 148 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0306] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 149. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 149 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0307] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 150. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 150 with 1, 2, 3, 4 or 5 conservative amino acid substitutions. In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 183. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 183 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0308] In some embodiments, the fusion protein comprises the amino acid sequence set forth 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 to the sequence set forth in SEQ ID NO: 184. In some embodiments, the fusion protein comprises the sequence set forth in SEQ ID NO: 184 with 1, 2, 3, 4 or 5 conservative amino acid substitutions.

[0309] In some embodiments, the fusion protein is a dimer comprising two fusion proteins described herein, wherein the two fusion proteins are the same. In some embodiments, the fusion protein is a dimer comprising two fusion proteins described herein, wherein the two fusion proteins are different. In some embodiments, the fusion protein is a dimer comprising a fusion protein described herein and a half-life extending domain lacking an IGFBPL1 polypeptide or fragment thereof. In some embodiments, the half-life extending domain of one or more fusion proteins comprises a modification to induce dimer formation (e.g., knob-in-hole modifications of Fc domains).

[0310] 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 to the sequence set forth 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 to the sequence set forth 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.

[0311] 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 to the sequence set forth 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 to the sequence set forth 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.

[0312] 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 to the sequence set forth 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 to the sequence set forth 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.

[0313] 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 to the sequence set forth 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 to the sequence set forth 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.

[0314] 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 to the sequence set forth 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 to the sequence set forth 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.

[0315] Methods of Making Polypeptides

[0316] Methods of making polypeptides are known in the art as described in WO 2016 / 025647 and provide below. In some embodiments, the polypeptides and fusion proteins described herein are made in transformed host cells using recombinant DNA techniques. To do so, a recombinant DNA molecule coding for the peptide is prepared. Methods of preparing such DNA molecules are well known in the art. For instance, sequences coding for the peptides could be excised from DNA using suitable restriction enzymes. Alternatively, the DNA molecule could be synthesized using chemical synthesis techniques, such as the phosphoramidate method. Also, a combination of these techniques could be used.

[0317] The methods of making polypeptides also include a vector capable of expressing the peptides in an appropriate host. The vector comprises the DNA molecule that codes for the peptides operatively linked to appropriate expression control sequences. Methods of affecting this operative linking, either before or after the DNA molecule is inserted into the vector, are well known. Expression control sequences include promoters, activators, enhancers, operators, ribosomal nuclease domains, start signals, stop signals, cap signals, polyadenylation signals, and other signals involved with the control of transcription or translation.

[0318] The resulting vector having the DNA molecule thereon is used to transform an appropriate host. This transformation may be performed using methods well known in the art.

[0319] Any of a large number of available and well-known host cells may be suitable for use in the methods disclosed herein. The selection of a particular host is dependent upon a number of factors recognized by the art. These include, for example, compatibility with the chosen expression vector, toxicity of the peptides encoded by the DNA molecule, rate of transformation, ease of recovery of the peptides, expression characteristics, bio-safety and costs. A balance of these factors must be struck with the understanding that not all hosts may be equally effective for the expression of a particular DNA sequence. Within these general guidelines, useful microbial hosts include bacteria (such as E. coli sp.), yeast (such as Saccharomyces sp.) and other fungi, insects, plants, mammalian (including human) cells in culture, or other hosts known in the art. Next, the transformed host is cultured and purified. Host cells may be cultured under conventional fermentation conditions so that the desired compounds are expressed. Such fermentation conditions are well known in the art. Finally, the peptides are purified from culture by methods well known in the art.

[0320] The compounds may also be made by synthetic methods. For example, solid phase synthesis techniques may be used. Suitable techniques are well known in the art, and include those described in Merrifield (1973), Chem. Polypeptides, pp. 335-61 (Katsoyannis and Panayotis eds.), Merrifield (1963), J. Am. Chem. Soc. 85: 2149; Davis et al. (1985), Biochem. Inti. 10: 394-414; Stewart and Young (1969), Solid Phase Peptide Synthesis; U.S. Pat. No. 3,941,763; Finn et al. (1976), The Proteins (3rd ed.) 2: 105-253; and Erickson et al. (1976), The Proteins (3rd ed.) 2: 257-527. Solid phase synthesis is the preferred technique of making individual peptides since it is the most cost-effective method of making small peptides.

[0321] Compounds that contain derivatized peptides or which contain non-peptide groups may be synthesized by well-known organic chemistry techniques.

[0322] Other methods are of molecule expression / synthesis are generally known in the art to one of ordinary skill.

[0323] The nucleic acid molecules described above can be contained within a vector that is capable of directing their expression in, for example, a cell that has been transduced with the vector. Accordingly, in addition to polypeptide mutants, expression vectors containing a nucleic acid molecule encoding a mutant and cells transfected with these vectors are among the certain embodiments.

[0324] Vectors suitable for use include T7 -based vectors for use in bacteria (see, for example, Rosenberg et al., Gene 56: 125, 1987), the pMSXND expression vector for use in mammalian cells (Lee and Nathans, J. Biol. Chem. 263:3521, 1988), and baculovirus-derived vectors (for example the expression vector pBacPAKS from Clontech, Palo Alto, Calif.) for use in insect cells. The nucleic acid inserts, which encode the polypeptide of interest in such vectors, can be operably linked to a promoter, which is selected based on, for example, the cell type in which expression is sought. For example, a T7 promoter can be used in bacteria, a polyhedrin promoter can be used in insect cells, and a cytomegalovirus or metal lothionein promoter can be used in mammalian cells. Also, in the case of higher eukaryotes, tissue-specific and cell type- specific promoters are widely available. These promoters are so named for their ability to direct expression of a nucleic acid molecule in a given tissue or cell type within the body. Skilled artisans are well aware of numerous promoters and other regulatory elements which can be used to direct expression of nucleic acids.

[0325] In addition to sequences that facilitate transcription of the inserted nucleic acid molecule, vectors can contain origins of replication, and other genes that encode a selectable marker. For example, the neomycin -resistance (neo1) gene imparts G418 resistance to cells in which it is expressed, and thus permits phenotypic selection of the transfected cells. Those of skill in the art can readily determine whether a given regulatory element or selectable marker is suitable for use in a particular experimental context.

[0326] Viral vectors that are suitable for use include, for example, retroviral, adenoviral, and adeno-associated vectors, herpes virus, simian virus 40 (SV40), and bovine papilloma virus vectors (see, for example, Gluzman (Ed.), Eukaryotic Viral Vectors, CSH Laboratory Press, Cold Spring Harbor, N.Y.).

[0327] Prokaryotic or eukaryotic cells that contain and express a nucleic acid molecule that encodes a polypeptide mutant are also suitable for use. A cell is a transfected cell, i.e., a cell into which a nucleic acid molecule, for example a nucleic acid molecule encoding a mutant polypeptide, has been introduced by means of recombinant DNA techniques. The progeny of such a cell are also considered suitable for use in the methods disclosed herein.

[0328] The precise components of the expression system are not critical. For example, a polypeptide mutant can be produced in a prokaryotic host, such as the bacterium E. coli, or in a eukaryotic host, such as an insect cell (e.g., an Sf21 cell), 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.). In selecting an expression system, it matters only that the components are compatible with one another. Artisans or ordinary skill are able to make such a determination. Furthermore, if guidance is required in selecting an expression system, skilled artisans may consult Ausubel et al. (Current Protocols in Molecular Biology, John Wiley and Sons, New York, N.Y., 1993) and Pouwels et al. (Cloning Vectors: A Laboratory Manual, 1985 Suppl. 1987).

[0329] The expressed polypeptides can be purified from the expression system using routine biochemical procedures, and can be used, e.g., as therapeutic agents, as described herein. Pharmaceutical Compositions

[0330] The disclosure provides pharmaceutical compositions of the IGFBPL1 polypeptides and fusion proteins described herein with a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant, a pharmaceutical composition comprising a cancer vaccine with a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant, a pharmaceutical composition comprising a therapeutic antibody with a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant, or a pharmaceutical composition comprising an immune checkpoint blocker with a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant.

[0331] In some embodiments, the pharmaceutical compositions comprise an IGFBPL1 polypeptide or fusion protein described herein, with a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative and / or adjuvant. In some embodiments, acceptable formulation materials preferably are nontoxic to recipients at the dosages and concentrations employed. In some embodiments, the formulation material(s) are for s.c. and / or I.V. administration. In certain embodiments, the pharmaceutical composition can contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In some embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite or sodium hydrogen- sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, betacyclodextrin or hydroxypropyl-beta- cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins), proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone), low molecular weight polypeptides, saltforming 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 glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate 80, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles, diluents, excipients and / or pharmaceutical adjuvants. (Remington's Pharmaceutical Sciences, 18th Edition, A. R. Gennaro, ed., Mack Publishing Company (1995). In certain embodiments, the formulation comprises PBS; 20 mMNaOAC, 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 one skilled in the art depending upon, for example, the intended route of administration, delivery format and desired dosage. See, for example. Remington's Pharmaceutical Sciences, supra. In some embodiments, such compositions may influence the physical state, stability, rate of in vivo release and rate of in vivo clearance of the IGFBPL1 polypeptide or fusion protein.

[0332] In some embodiments, the primary vehicle or carrier in a pharmaceutical composition can be either aqueous or non-aqueous in nature. For example, in some embodiments, a suitable vehicle or carrier can 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 comprises isotonic phosphate- buffered saline. In some embodiments, neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In some embodiments, pharmaceutical compositions comprise Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, which can further include sorbitol or a suitable substitute therefore. In some embodiments, a composition comprising a fusion protein can be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (Remington's Pharmaceutical Sciences, supra) in the form of a lyophilized cake or an aqueous solution. Further, in some embodiments, a composition comprising a fusion protein is formulated as a lyophilizate using appropriate excipients such as sucrose.

[0333] In some embodiments, the pharmaceutical composition can be selected for parenteral delivery. In some embodiments, the compositions can be selected for inhalation or for delivery' through the digestive tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the ability of one skilled in the art. 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 administration to the eye.

[0334] In some embodiments, the formulation components are present in concentrations that are acceptable to the site of administration. In some embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8.

[0335] In some embodiments, when parenteral administration is contemplated, a therapeutic composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution comprising the fusion protein, in a pharmaceutically acceptable vehicle. In some embodiments, a vehicle for parenteral injection is sterile distilled water in which the fusion protein is formulated as a sterile, isotonic solution, properly preserved. In certain embodiments, the preparation can involve the formulation of the desired molecule with an agent, such as injectable microspheres, bio-erodible particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads or liposomes, that can provide for the controlled or sustained release of the product which can then be delivered via a depot injection. In certain embodiments, hyaluronic acid can also be used, and can have the effect of promoting sustained duration in the circulation. In certain embodiments, implantable drug delivery devices can be used to introduce the desired molecule.

[0336] In some embodiments, a 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 comprising the fusion protein is formulated with a propellant for aerosol delivery' . In certain embodiments, solutions can be nebulized. Pulmonary administration is further described in PCT application No. PCT / US94 / 00I875, which describes pulmonary' delivery / of chemically modified proteins.

[0337] In some embodiments, it is contemplated that formulations can be administered orally. In some embodiments, the fusion protein that is administered in this fashion can be formulated with or without those carriers customarily used in the compounding of solid dosage forms such as tablets and capsules. In some embodiments, a capsule can be designed to release the active portion of the formulation at the point in the gastrointestinal tract when bioavailability is maximized and pre-systemic degradation is minimized. In some embodiments, at least one additional agent is included to facilitate absorption of the fusion protein. In some embodiments, diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binders can also be employed.

[0338] In some embodiments, a pharmaceutical composition includes an effective quantity of the fusion protein in a mixture with non-toxic excipients which are suitable for the manufacture of tablets. In certain embodiments, by dissolving the tablets in sterile water, or another appropriate vehicle, solutions can be prepared in unit-dose form. In some embodiments, suitable excipients include, but are not limited to, inert diluents, such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate; or binding agents, such as starch, gelatin, or acacia; or lubricating agents such as magnesium stearate, stearic acid, or talc.

[0339] Additional pharmaceutical compositions will be evident to those skilled in the art, including formulations involving the fusion protein in sustained- or controlled-delivery formulations. In some embodiments, techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. See for example, PCT Application No. PCT / US93 / 00829 which describes the controlled release of porous polymeric microparticles for the delivery of pharmaceutical compositions. In some embodiments, sustained-release preparations can include semipermeable polymer matrices in the form of shaped articles, e.g. films, or microcapsules. Sustained release matrices can include polyesters, hydrogels, polylactides (U.S. Pat. No. 3,773,919 and EP 058,481), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al.. Biopolymers, 22:547-556 (1983)), poly (2- hydroxyethyl-methaciylate) (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., supra) or poly- D(-)-3 -hydroxybutyric acid (EP 133,988). In certain embodiments, sustained release compositions can also include liposomes, which can be prepared by any of several methods known in the art. See, e.g., Eppstein et al, Proc. Natl. Acad. Sci. USA, 82:3688-3692 (1985); EP 036,676, EP 088,046 and EP 143,949.

[0340] The pharmaceutical composition to be used for in vivo administration typically is sterile. In certain embodiments, this can be accomplished by filtration through sterile filtration membranes. In certain embodiments, where the composition is lyophilized, sterilization using this method can be conducted either prior to or following lyophilization and reconstitution. In certain embodiments, the composition for parenteral administration can be stored in lyophilized form or in a solution. In certain embodiments, parenteral compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0341] In some embodiments, once the pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or as a dehydrated or lyophilized powder. In certain embodiments, such formulations can be stored either in a ready- to-use form or in a form (e.g., lyophilized) that is reconstituted prior to administration.

[0342] In some embodiments, kits are provided for producing a single-dose administration unit. In certain embodiments, the kit can contain both a first container having a dried protein and a second container having an aqueous formulation. In certain embodiments, kits containing single and multi-chambered pre-filled syringes (e.g., liquid syringes and lyosyringes) are included.

[0343] In some embodiments, the effective amount of a pharmaceutical composition comprising the fusion protein to be employed therapeutically will depend, for example, upon the therapeutic context and objectives. One skilled in the art will appreciate that the appropriate dosage levels for treatment, according to certain embodiments, will thus vary' depending, in part, upon the molecule delivered, the indication for which the fusion protein is being used, the route of administration, and the size (body weight, body surface or organ size) and / or condition (the age and general health) of the patient. In some embodiments, the clinician can titer the dosage and modify the route of administration to obtain the optimal therapeutic effect.

[0344] In some embodiments, the frequency of dosing will take into account the pharmacokinetic parameters of the fusion protein in the formulation used. In some embodiments, a clinician will administer the composition until a dosage is reached that achieves the desired effect. In certain embodiments, the composition can 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 implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. In some embodiments, appropriate dosages can be ascertained through use of appropriate dose-response data. In some embodiments, the route of administration of the pharmaceutical composition is in accord with known methods, e.g. orally, through injection by intravenous, intraperitoneal, intracerebral (intra-parenchymal), intracerebroventricular, intramuscular, subcutaneously, intraocular, intraarterial, intraportal, or intralesional routes; by sustained release systems or by implantation devices. In certain embodiments, the compositions can be administered by bolus injection or continuously by infusion, or by implantation device. In some embodiments, individual elements of the combination therapy may be administered by different routes.

[0345] In some embodiments, the composition is administered locally via implantation of a membrane, sponge or another appropriate material onto which the desired molecule has been absorbed or encapsulated. In some embodiments, where an implantation device is used, the device can be implanted into any suitable tissue or organ, and delivery of the desired molecule can be via diffusion, timed-release bolus, or continuous administration. In some embodiments, it can be desirable to use a pharmaceutical composition the fusion protein in an ex vivo manner. In such instances, cells, tissues and / or organs that have been removed from the patient are exposed to a pharmaceutical composition comprising the fusion protein after which the cells, tissues and / or organs are subsequently implanted back into the patient.

[0346] Kits

[0347] In some embodiments, the disclosure provides a kit comprising the IGFBPL1 polypeptide or IGFBPL1 fragment described herein and instructions for use. In some embodiments, the disclosure provides a kit comprising the fusion protein and instructions for use. The kits may comprise, in a suitable container, the fusion protein, one or more controls, and various buffers, reagents, enzymes and other standard ingredients well known in the art. The container can include at least one vial, well, test tube, flask, bottle, syringe, or other container means, into which the fusion protein may be placed, and in some instances, suitably aliquoted. Where an additional component is provided, the kit can contain additional containers into which this component may be placed. The kits can also include a means for containing the fusion protein, and any other reagent containers in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained. Containers and / or kits can include labeling o with instructions for use and / or warnin ogs. Methods of Use

[0348] In some embodiments, the disclosure provides methods of using the IGFBPL1 polypeptides described herein. In some embodiments, the disclosure provides methods of using the IGFBPL1 fragments described herein. In some embodiments, the disclosure provides methods of using the fusion proteins described herein. In any of the methods described herein, the fusion protein is monovalent, bivalent, trivalent or tetravalent.

[0349] In some embodiments, the disclosure provides a method of inhibiting SORT1 activity in a cell comprising contacting the cell with an IGFBPL1 polypeptide or an IGFBPL1 fragment described herein. In some embodiments, the disclosure provides a method of inhibiting SORT1 activity in a cell comprising contacting the cell with a fusion protein described herein. In some embodiments, the cell is a microglia. In some embodiments, the microglia is in the brain or eye of a subject. In some embodiments, the disclosure provides a method of increasing pro- inflammatory cytokine secretion from a cell by contacting the cell with a fusion protein described herein. In some embodiments, the cell is a microglia. In some embodiments, the cytokine is IL- 10, GM-CSF, MIP-1 alpha, or any combination thereof.

[0350] In some embodiments, the disclosure provides a method of inhibiting inflammatory cytokine secretion from a cell by contacting the cell with an IGFBPL1 polypeptide or an IGFBPL1 fragment described herein. In some embodiments, the disclosure provides a method of inhibiting inflammatory cytokine secretion from a cell by contacting the cell with a fusion protein described herein. In some embodiments, the cell is a microglia. In some embodiments, the cytokine is IL-6.

[0351] Methods of quantifying protein are known in the art and include, for example and without limitation, ELISA and Western blot.

[0352] In some embodiments, the disclosure provides a method of inhibiting DCC activity in a cell comprising contacting the cell with an IGFBPL1 polypeptide or an IGFBPL1 fragment described herein. In some embodiments, the disclosure provides a method of inhibiting DCC activity in a cell comprising contacting the cell with a fusion protein described herein. In some embodiments, the cell is a neuron. In some embodiments, the neuron is in the brain or eye of a subject. In some embodiments, the disclosure provides a method of increasing the number of neurons and / or neurite outgrowth comprising contacting the neuron with a fusion protein described herein. Methods for determining the number of neurons are known to those of skill in the art. For example, a population of cells comprising neurons can be manually counted. Methods for determining neurite outgrowth are known to those of skill in the art. For example, a population of cells comprising neurons can be fixed and imaged to visualize neurite extension. Measurements of neurite extension can be manually determined.

[0353] In some embodiments, the disclosure provides a method for inducing phagocytic activity of microglia by contacting the microglia with an IGFBPL1 polypeptide or an IGFBPL1 fragment described herein. In some embodiments, the disclosure provides a method for inducing phagocytic activity of microglia by contacting the microglia with a fusion protein described herein. Methods for determining phagocytic activity are known to those of skill in the art and include use of fluorescent latex beads or particles, such as Zymosan, and subsequently counting the number of cells that ingest the beads, wherein an increase in bead ingestion indicates an increase in phagocytic activity.

[0354] The IGFBPL1 polypeptides or IGFBPL1 fragments described herein may also be used for preferential targeting of neurons over microglia or vice versa. The fusion proteins described herein may also be used for preferential targeting of neurons over microglia or vice versa. Without wishing to be bound by theory, it is believed that the two ends of hIGFBPLI are differentially involved in mediating cell binding such that both ends of IGFBPL1 are important for mediating neuroprotective (C -terminus) and anti-inflammatory (N-terminus) activities. This is believed to be due to the N-terminal domain of IGFBPL1 binding to microglia and the C- terminal domain of IGFBPL1 binding to neurons.

[0355] An IGFBPL1 polypeptide or an IGFBPL1 fragment that exhibits “preferential targeting” of neurons over microglia shows stronger binding to neurons than microglia. In some embodiments, the IGFBPL1 polypeptide or the IGFBPL1 fragment preferentially targeting neurons binds to neurons with an affinity that is at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 25 times higher than the affinity with which the IGFBPL1 polypeptide or the IGFBPL1 fragment binds to microglia.

[0356] A fusion protein or fusion protein dimer that exhibits “preferential targeting” of neurons over microglia shows stronger binding to neurons than microglia. In some embodiments, the fusion protein or fusion protein dimer preferentially targeting neurons binds to neurons with an affinity that is at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 25 times higher than the affinity with which the fusion protein or fusion protein dimer binds to microglia.

[0357] An IGBFPL1 polypeptide or an IGFBPL1 fragment that exhibits “preferential targeting” of microglia over neurons shows stronger binding to microglia than neurons. In some embodiments, the IGFBPL1 polypeptide or the IGFBPL1 fragment preferentially targeting microglia binds to microglia with an affinity that is at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 25 times higher than the affinity with which the IGFBPL1 polypeptide or the IGFBPL1 fragment binds to neurons.

[0358] A fusion protein or fusion protein dimer that exhibits “preferential targeting” of microglia over neurons shows stronger binding to microglia than neurons. In some embodiments, the fusion protein or fusion protein dimer preferentially targeting microglia binds to microglia with an affinity that is at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 25 times higher than the affinity with which the fusion protein or fusion protein dimer binds to neurons.

[0359] Thus provided herein is a method for preferential targeting of neurons over microglia in a sample or subject, comprising introducing to the sample or subject an IGFBPL1 fragment comprising the C-terminal domain of IGFBPL1. Also provided herein is a method for preferential targeting of neurons over microglia in a sample or subject, comprising introducing to the sample or subject an N-terminal fusion protein dimer described herein. In some embodiments, fusion proteins that preferentially bind DCC relative to SORT1 are used for preferential targeting of neurons over microglia

[0360] Thus provided herein is a method for preferential targeting of microglia over neurons in a sample or subject, comprising introducing to the sample or subject an IGFBPL1 fragment comprising the N-terminal domain of IGFBPL1. Also provided herein is a method for preferential targeting of microglia over neurons in a sample or subject, comprising introducing to the sample or subject an C-terminal fusion protein dimer described herein. Methods of Treatment

[0361] In some aspects, the disclosure provides methods for treating a subject with a disease or disorder with the IGFBPL1 polypeptides, IGFBPL1 fragments, fusion proteins, or compositions described herein.

[0362] In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein. In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 fragment or composition thereof described herein. In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject a fusion protein or composition thereof described herein.

[0363] In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein, wherein the IGFBPL1 polypeptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 151-157. In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein, wherein the IGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID Nos: 151-157.

[0364] In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179. In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179.

[0365] In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179, provided no other domains of an IGFBPL1 polypeptide are present in the IGFBPL1 fragment. In some aspects, the disclosure provides a method for treating a subject with a retinal degenerative disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179, provided no other domains of an IGFBPL1 polypeptide are present in the IGFBPL1 fragment.

[0366] In some aspects, the 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 to an amino acid sequence selected from SEQ ID Nos: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184. In some aspects, the 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 Nos: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184.

[0367] In some aspects, the 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 to SEQ ID NO: 15. In some aspects, the 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.

[0368] In some aspects, the 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 to SEQ ID NO: 16. In some aspects, the 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.

[0369] In some aspects, the 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 to SEQ ID NO: 17. In some aspects, the 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:

[0370] 17.

[0371] In some aspects, the 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 to SEQ ID NO: 18. In some aspects, the 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:

[0372] 18.

[0373] In some aspects, the 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 to SEQ ID NO: 19. In some aspects, the 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:

[0374] 19.

[0375] In some aspects, the 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 to SEQ ID NO: 20. In some aspects, the 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:

[0376] 20.

[0377] In some aspects, the 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 to SEQ ID NO: 21. In some aspects, the 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:

[0378] 21.

[0379] In some aspects, the 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 to SEQ ID NO: 22. In some aspects, the 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:

[0380] 22.

[0381] In some aspects, the 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 to SEQ ID NO: 51. In some aspects, the 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.

[0382] In some aspects, the 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 to SEQ ID NO: 52. In some aspects, the 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:

[0383] 53.

[0384] In some aspects, the 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 to SEQ ID NO: 54. In some aspects, the 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:

[0385] 54.

[0386] In some aspects, the 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 to SEQ ID NO: 55. In some aspects, the 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:

[0387] 55.

[0388] In some aspects, the 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 to SEQ ID NO: 56. In some aspects, the 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:

[0389] 56.

[0390] In some aspects, the 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 to SEQ ID NO: 57. In some aspects, the 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:

[0391] 57.

[0392] In some aspects, the 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 to SEQ ID NO: 58. In some aspects, the 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:

[0393] 58.

[0394] In some aspects, the 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 to SEQ ID NO: 59. In some aspects, the 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:

[0395] 59.

[0396] In some aspects, the 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 to SEQ ID NO: 60. In some aspects, the 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:

[0397] 60. In some aspects, the 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 to SEQ ID NO: 61. In some aspects, the 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:

[0398] 61.

[0399] In some aspects, the 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 to SEQ ID NO: 62. In some aspects, the 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:

[0400] 62.

[0401] In some aspects, the 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 to SEQ ID NO: 63. In some aspects, the 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:

[0402] 63.

[0403] In some aspects, the 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 to SEQ ID NO: 64. In some aspects, the 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.

[0404] In some aspects, the 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 to SEQ ID NO: 65. In some aspects, the 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:

[0405] 65.

[0406] In some aspects, the 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 to SEQ ID NO: 66. In some aspects, the 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:

[0407] 66.

[0408] In some aspects, the 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 to SEQ ID NO: 67. In some aspects, the 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:

[0409] 67.

[0410] In some aspects, the 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 to SEQ ID NO: 68. In some aspects, the 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.

[0411] In some aspects, the 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 to SEQ ID NO: 69. In some aspects, the 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:

[0412] 69.

[0413] In some aspects, the 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 to SEQ ID NO: 70. In some aspects, the 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:

[0414] 70.

[0415] In some aspects, the 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 to SEQ ID NO: 71. In some aspects, the 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:

[0416] 71.

[0417] In some aspects, the 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 to SEQ ID NO: 72. In some aspects, the 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:

[0418] 72.

[0419] In some aspects, the 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 to SEQ ID NO: 73. In some aspects, the 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:

[0420] 73.

[0421] In some aspects, the 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 to SEQ ID NO: 74. In some aspects, the 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:

[0422] 74.

[0423] In some aspects, the 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 to SEQ ID NO: 75. In some aspects, the 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:

[0424] 75.

[0425] In some aspects, the 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 to SEQ ID NO: 76. In some aspects, the 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:

[0426] 76.

[0427] In some aspects, the 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 to SEQ ID NO: 77. In some aspects, the 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:

[0428] 77.

[0429] In some aspects, the 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 to SEQ ID NO: 78. In some aspects, the 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:

[0430] 78.

[0431] In some aspects, the 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 to SEQ ID NO: 79. In some aspects, the 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:

[0432] 79.

[0433] In some aspects, the 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 to SEQ ID NO: 80. In some aspects, the 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:

[0434] 80.

[0435] In some aspects, the 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 to SEQ ID NO: 81. In some aspects, the 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:

[0436] 81.

[0437] In some aspects, the 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 to SEQ ID NO: 82. In some aspects, the 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:

[0438] 82.

[0439] In some aspects, the 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 to SEQ ID NO: 83. In some aspects, the 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:

[0440] 83. In some aspects, the 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 to SEQ ID NO: 84. In some aspects, the 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:

[0441] 84.

[0442] In some aspects, the 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 to SEQ ID NO: 85. In some aspects, the 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:

[0443] 85.

[0444] In some aspects, the 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 to SEQ ID NO: 86. In some aspects, the 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:

[0445] 86.

[0446] In some aspects, the 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 to SEQ ID NO: 94. In some aspects, the 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.

[0447] In some aspects, the 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 to SEQ ID NO: 95. In some aspects, the 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:

[0448] 95.

[0449] In some aspects, the 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 to SEQ ID NO: 96. In some aspects, the 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:

[0450] 96.

[0451] In some aspects, the 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 to SEQ ID NO: 97. In some aspects, the 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:

[0452] 97.

[0453] In some aspects, the 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 to SEQ ID NO: 98. In some aspects, the 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.

[0454] In some aspects, the 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 to SEQ ID NO: 99. In some aspects, the 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:

[0455] 99.

[0456] In some aspects, the 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 to SEQ ID NO: 100. In some aspects, the 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:

[0457] 100.

[0458] In some aspects, the 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 to SEQ ID NO: 101. In some aspects, the 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:

[0459] 101.

[0460] In some aspects, the 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 to SEQ ID NO: 102. In some aspects, the 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:

[0461] 102.

[0462] In some aspects, the 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 to SEQ ID NO: 103. In some aspects, the 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:

[0463] 103.

[0464] In some aspects, the 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 to SEQ ID NO: 104. In some aspects, the 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:

[0465] 104.

[0466] In some aspects, the 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 to SEQ ID NO: 105. In some aspects, the 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:

[0467] 105.

[0468] In some aspects, the 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 to SEQ ID NO: 106. In some aspects, the 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:

[0469] 106.

[0470] In some aspects, the 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 to SEQ ID NO: 107. In some aspects, the 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:

[0471] 107.

[0472] In some aspects, the 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 to SEQ ID NO: 108. In some aspects, the 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:

[0473] 108.

[0474] In some aspects, the 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 to SEQ ID NO: 109. In some aspects, the 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:

[0475] 109.

[0476] In some aspects, the 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 to SEQ ID NO: 110. In some aspects, the 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:

[0477] 110.

[0478] In some aspects, the 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 to SEQ ID NO: 111. In some aspects, the 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:

[0479] 111.

[0480] In some aspects, the 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 to SEQ ID NO: 112. In some aspects, the 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:

[0481] 112.

[0482] In some aspects, the 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 to SEQ ID NO: 113. In some aspects, the 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:

[0483] 113. In some aspects, the 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 to SEQ ID NO: 114. In some aspects, the 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:

[0484] 114.

[0485] In some aspects, the 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 to SEQ ID NO: 115. In some aspects, the 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:

[0486] 115.

[0487] In some aspects, the 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 to SEQ ID NO: 116. In some aspects, the 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:

[0488] 116.

[0489] In some aspects, the 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 to SEQ ID NO: 117. In some aspects, the 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.

[0490] In some aspects, the 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 to SEQ ID NO: 118. In some aspects, the 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:

[0491] 118.

[0492] In some aspects, the 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 to SEQ ID NO: 119. In some aspects, the 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:

[0493] 119.

[0494] In some aspects, the 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 to SEQ ID NO: 120. In some aspects, the 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:

[0495] 120.

[0496] In some aspects, the 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 to SEQ ID NO: 121. In some aspects, the 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.

[0497] In some aspects, the 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 to SEQ ID NO: 122. In some aspects, the 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:

[0498] 122.

[0499] In some aspects, the 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 to SEQ ID NO: 123. In some aspects, the 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:

[0500] 123.

[0501] In some aspects, the 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 to SEQ ID NO: 124. In some aspects, the 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:

[0502] 124.

[0503] In some aspects, the 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 to SEQ ID NO: 125. In some aspects, the 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:

[0504] 125.

[0505] In some aspects, the 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 to SEQ ID NO: 126. In some aspects, the 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:

[0506] 126.

[0507] In some aspects, the 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 to SEQ ID NO: 127. In some aspects, the 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:

[0508] 127.

[0509] In some aspects, the 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 to SEQ ID NO: 128. In some aspects, the 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:

[0510] 128.

[0511] In some aspects, the 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 to SEQ ID NO: 129. In some aspects, the 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:

[0512] 129.

[0513] In some aspects, the 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 to SEQ ID NO: 130. In some aspects, the 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:

[0514] 130.

[0515] In some aspects, the 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 to SEQ ID NO: 131. In some aspects, the 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:

[0516] 131.

[0517] In some aspects, the 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 to SEQ ID NO: 132. In some aspects, the 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:

[0518] 132.

[0519] In some aspects, the 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 to SEQ ID NO: 133. In some aspects, the 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:

[0520] 133.

[0521] In some aspects, the 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 to SEQ ID NO: 134. In some aspects, the 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:

[0522] 134.

[0523] In some aspects, the 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 to SEQ ID NO: 135. In some aspects, the 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:

[0524] 135.

[0525] In some aspects, the 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 to SEQ ID NO: 136. In some aspects, the 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:

[0526] 136. In some aspects, the 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 to SEQ ID NO: 138. In some aspects, the 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:

[0527] 138.

[0528] In some aspects, the 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 to SEQ ID NO: 139. In some aspects, the 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:

[0529] 139.

[0530] In some aspects, the 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 to SEQ ID NO: 140. In some aspects, the 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:

[0531] 140.

[0532] In some aspects, the 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 to SEQ ID NO: 142. In some aspects, the 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.

[0533] In some aspects, the 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 to SEQ ID NO: 144. In some aspects, the 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:

[0534] 144.

[0535] In some aspects, the 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 to SEQ ID NO: 145. In some aspects, the 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:

[0536] 145.

[0537] In some aspects, the 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 to SEQ ID NO: 146. In some aspects, the 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:

[0538] 146.

[0539] In some aspects, the 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 to SEQ ID NO: 147. In some aspects, the 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.

[0540] In some aspects, the 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 to SEQ ID NO: 148. In some aspects, the 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:

[0541] 148.

[0542] In some aspects, the 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 to SEQ ID NO: 149. In some aspects, the 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:

[0543] 149.

[0544] In some aspects, the 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 to SEQ ID NO: 150. In some aspects, the 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:

[0545] 150.

[0546] In some aspects, the 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 to SEQ ID NO: 183. In some aspects, the 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:

[0547] 183.

[0548] In some aspects, the 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 to SEQ ID NO: 184. In some aspects, the 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:

[0549] 184.

[0550] In some aspects, the 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 to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 21. In some aspects, the 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.

[0551] In some aspects, the 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 to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 9. 1 In some aspects, the 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.

[0552] In some aspects, the 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 to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 21. In some aspects, the 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.

[0553] In some aspects, the 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 to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 139. In some aspects, the 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.

[0554] In some aspects, the 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 to the sequence set forth 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

[0555] I l l 97%, at least 98%, or at least 99% identity to the sequence set forth in SEQ ID NO: 141. In some aspects, the 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.

[0556] In some aspects, the 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 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 Stargardt 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 Stargardt disease.

[0557] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 fragment or composition thereof described herein. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject a fusion protein or composition thereof described herein.

[0558] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein, wherein the IGFBPL1 polypeptide comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 151-157. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 polypeptide or composition thereof described herein, wherein the IGFBPL1 polypeptide comprises an amino acid sequence selected from SEQ ID Nos: 151-157.

[0559] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179.

[0560] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179, provided no other domains of an IGFBPL1 polypeptide are present in the IGFBPL1 fragment. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory disease, comprising administering to the subject an IGFBPL1 fragment or composition described herein, wherein the IGFBPL1 fragment comprises an amino acid sequence selected from SEQ ID Nos: 2-3, 25-28, 37-50, 158, and 160-179, provided no other domains of an IGFBPL1 polypeptide are present in the IGFBPL1 fragment.

[0561] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to an amino acid sequence selected from SEQ ID Nos: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 Nos: 15-22, 51-86, 94-136, 138-140, 142, 144-150, and 183-184. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 15. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0562] 15.

[0563] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 16. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0564] 16.

[0565] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 17. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0566] 17.

[0567] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 18. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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.

[0568] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 19. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0569] 19.

[0570] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 20. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0571] 20.

[0572] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 21. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0573] 21.

[0574] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 22. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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.

[0575] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 51. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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.

[0576] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 52. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0577] 53.

[0578] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 54. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0579] 54.

[0580] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 55. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0581] 55.

[0582] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 56. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0583] 56.

[0584] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 57. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0585] 57.

[0586] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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 to SEQ ID NO: 58. In some aspects, the disclosure provides a method for treating a subject with a retinal inflammatory 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:

[0587] 58.

[0588] In some aspects, the disclosure provides a method for treating a subject with a retinal inflammat...

Claims

CLAIMS1. A fusion protein comprising a half-life extending domain and a human IGFBPL1 (hIGFBPLI) polypeptide or a fragment thereof, wherein the half-life extending domain is operatively linked to the hIGFBPLI polypeptide or fragment thereof with or without a linker.

2. The fusion protein of claim 1, wherein the hIGFBPLI polypeptide or the fragment thereof comprises a thumb domain, an N-terminal domain, a Kazal domain, and 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 of any one of claims 1-3, wherein the fragment of hIGFBPLI does not comprise the thumb domain of hIGFBPLI.

5. The fusion protein of any one of claims 1-4, wherein the fragment of hIGFBPLI does not comprise the N-terminal domain of hIGFBPLI.

6. The fusion protein of any one of claims 1-5, wherein the fragment of hIGFBPLI does not comprise the Kazal domain of hIGFBPLI.

7. The fusion protein of any one of claims 1-6, wherein the fragment of hIGFBPLI does not comprise the Ig-like domain of hIGFBPLI.

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

9. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the N-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

10. The fusion protein of claim 9, wherein the fragment of hIGFBPLI comprises the amino acid sequence selected from SEQ ID NOs: 2, 158, and 160-166.

11. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

12. The fusion protein of claim 11, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 179.

13. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the thumb domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

14. The fusion protein of claim 13, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 25.

15. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

16. The fusion protein of claim 15, wherein the fragment of hIGFBPLI comprises the amino acid sequence selected from SEQ ID Nos: 26, 167, and 169.

17. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the Kazal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

18. The fusion protein of claim 17, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 27 or SEQ ID NO: 178.

19. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the Ig-like domain and C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

20. The fusion protein of claim 19, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 168 or SEQ ID NO: 170.

21. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the Kazal domain and the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

22. The fusion protein of claim 21, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 171 or SEQ ID NO: 173.

23. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the Kazal domain, the Ig-like domain, and the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

24. The fusion protein of claim 23, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 172 or SEQ ID NO: 174.

25. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the N-terminal domain and the Ig-like domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

26. The fusion protein of claim 25, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 175.

27. The fusion protein of any one of claims 1-3, wherein the fragment of hIGFBPLI comprises the N-terminal domain, the Ig-like domain, and the C-terminal domain of hIGFBPLI, provided no other domain of hIGFBPLI is part of the hIGFBPLI fragment.

28. The fusion protein of claim 27, wherein the fragment of hIGFBPLI comprises the amino acid sequence of SEQ ID NO: 176 or SEQ ID NO: 177.

29. The fusion protein of any one of claims 1-24, wherein the hIGFBPLI polypeptide or the fragment of hIGFBPLI comprises one or more domains of murine IGFBPL1 (mIGFBPLI).

30. The fusion protein of claim 29, wherein the hIGFBPLI polypeptide or the fragment of hIGFBPLI comprises an amino acid sequence selected from SEQ ID Nos: 151-153 and 155-157.

31. The fusion protein of any one of claims 1-30, wherein the hIGFBPLI polypeptide or the fragment thereof comprises the native signal peptide of hIGFBPLI.

32. The fusion protein of claim 31, wherein the native signal peptide comprises the amino acid sequence of SEQ ID NO: 28.

33. The fusion protein of any one of claims 1-30, wherein the hIGFBPLI polypeptide or the 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 1-3 and 31-34, wherein the hIGFBPLI polypeptide comprises an amino acid sequence selected from SEQ ID NOs: 1, 10 and 11, or wherein the fragment of hIGFBPLI comprises an amino acid sequence selected from SEQ ID NOs: 37-50, 158 and 160-180.

36. The fusion protein of any one of claims 1-35, wherein the half-life extending domain is located at the N-terminus of the hIGFBPLI polypeptide or the fragment thereof.

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

38. The fusion protein of any one of claims 1-37, wherein the half-life extending domain is selected from an Fc domain, an albumin domain, a transferrin domain, a C-terminal peptide, an elastin-like peptide, a non-exact repeat peptide sequence, a proline-alanine-serine polymer, a repeat sequence of glycine rich polypeptides, and a gelatin-like protein.

39. The fusion protein of any one of claims 1-37, wherein the half-life extending 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 NOs: 4-9.

41. The fusion protein of any one of claims 1-37, wherein the half-life extending 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 1-42, wherein the half-life extending domain and the hIGFBPLI polypeptide or the fragment thereof are operably linked without a linker.

44. The fusion protein of any one of claims 1-42, wherein the half-life extending domain and the hIGFBPLI polypeptide or the fragment thereof are operably linked with 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 linker is a TEV linker or a GS linker.

47. The fusion protein of any one of claims 44-46, wherein the linker comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 12-14, 23-24, 159, and 182.

48. The fusion protein of claim 1, wherein the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 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 NOs: 15-22, 109-110, 134, 138-140, 142, 148-150.

50. The fusion protein of claim 1, wherein the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 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 NOs: 51-72, 111-133 and 183.

52. The fusion protein of claim 1, wherein the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID NOs: 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 NOs: 94-100, and 102-108.

54. The fusion protein of claim 1, wherein the fusion protein comprises an amino acid sequence that is 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% identical to an amino acid sequence selected from SEQ ID Nos: 144-147.

55. The fusion protein of claim 54, wherein the fusion protein comprises an amino acid sequence selected from SEQ ID NOs: 144-147.

56. The fusion protein of any one of claims 1-55, wherein the hIGFBPLI or the fragment thereof comprises at least one glycosylation mutation.

57. The fusion protein of claim 56, wherein the glycosylation mutation is at amino acid residue 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 hIGFBPLI polypeptide comprising the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NOs: 87-91.

60. The fusion protein of any one of claims 38-59, 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 NOs: 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 the glycosylation mutation comprises an amino acid sequence selected from SEQ ID NOs: 92 and 93.

63. A fusion protein dimer comprising two fusion proteins of any one of claims 1-62.

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

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 extending domain operably linked to a first hIGFBPLI polypeptide or a fragment thereof with or without a linker, and (ii) a second fusion protein comprising a second half-life extending domain operably linked to a second hIGFBPLI polypeptide or a fragment thereof with or without a linker.

67. The fusion protein dimer of claim 66, where the first and second IGFBPL1 polypeptide or the fragment thereof is the same.

68. The fusion protein dimer of claim 66, where the first and second IGFBPL1 polypeptide or the fragment thereof is different.

69. The fusion protein dimer of any one of claims 66-68, wherein the first and second halflife extending domain is the same.

70. The fusion protein dimer of any one of claims 66-68, wherein the first and second halflife extending domain is different.

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

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

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

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

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

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

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

78. The fusion protein dimer of any one of claims 75-77, wherein the third hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the half-life extending domain than the first hIGFBPLI polypeptide or the fragment thereof.

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

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

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

82. The fusion protein dimer of any one of claims 79-81, wherein the fourth hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the half-life extending domain than the second hIGFBPLI polypeptide or the fragment thereof.

83. The fusion protein dimer of any one of claims 66-82, wherein the first, second, third and / or fourth hIGFBPLI polypeptide or the fragment thereof, comprises an amino acid sequence selected from SEQ ID NOs: 1, 10-11, 37-50, 158 and 160-180.

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

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

86. The fusion protein dimer of claim 85, wherein the first Fc domain comprises knob mutations and the second Fc domain comprises hole mutations to dimerize the first and second Fc domains.

87. The fusion protein dimer of claim 85, wherein the first Fc domain comprises hole mutations and the second Fc domain comprises knob mutations to dimerize the first and second Fc chains.

88. The fusion protein dimer of any one of claims 84-86, wherein the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NOs: 4-9, 92 and 93.

89. The fusion protein dimer of any one of claims 66-88, 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) 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 comprisesthe 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: 140, and the second fusion protein comprises the amino acid sequence of SEQ ID NO: 141.

90. A fusion protein dimer comprising the fusion protein of any one of claims 1-62, and a second half-life extending domain, wherein the second half-life extending domain does not comprise an hIGFBPLI polypeptide or a fragment thereof.

91. The fusion protein dimer of claim 90, wherein the first and second half-life extending domain is the same.

92. The fusion protein dimer of claim 90, wherein the first and second half-life extending domain is different.

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

94. The fusion protein dimer of claim 93, wherein the first Fc domain comprises knob mutations and the second Fc domain comprises hole mutations to dimerize the first and second Fc domains.

95. The fusion protein dimer of claim 93, wherein the first Fc domain comprises hole mutations and the second Fc domain comprises knob mutations to dimerize the first and second Fc domains.

96. The fusion protein dimer of any one of claims 93-95, wherein the first and second Fc domains each comprise an amino acid sequence selected from SEQ ID NOs: 4-9, 92 and 93.

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

98. The fusion protein dimer of claim 97, wherein the second hIGFBPLI polypeptide or the fragment thereof is operably linked to the opposite end of the hIGFBPLI polypeptide or the 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 the first and second fusion proteins each comprise an amino acid sequence selected from SEQ ID NOs: 51-86, wherein the first and second fusion proteins are the same or different.

101. A composition comprising the fusion protein or fusion protein dimer of any one of claims 1-100.

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

103. The method of claim 102, wherein the fusion protein or fusion protein dimer binds DCC in a 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 average neurite length, number of neurites, number of neurons, and / or number of neurites per neuron.

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

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

108. The method of claim 107, wherein the fusion protein or fusion protein dimer binds SORT1 in a 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 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-1 alpha, and any combination thereof.

112. The method of any one of claims 109-111, wherein the fusion protein or fusion protein dimer inhibits production of one or more proteins by the microglia.

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 112-114, wherein the one or more proteins comprise IP- 10.

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

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

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

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

120. A method of treating a retinal degenerative disease in a subject, comprising administering to the subject the fusion protein or fusion protein dimer of any one of claims 1-100, or the composition of 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 Stargardt disease.

122. A method of treating a retinal inflammatory disease in a subject, comprising administering to the subject the fusion protein or fusion protein dimer of any one of claims 1- 100, or the composition of 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 120-123, wherein the fusion protein, fusion protein dimer, or composition is administered to the eye of the subject.

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

126. The method of claim 125, wherein local ocular administration comprises periocular, intravitreal, or intracameral administration.

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

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

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

130. The method of any one of claims 127-129, 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 Schilder’s disease.

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