Minibinder heterofusion proteins induce transdifferentiation

Abstract

Fusion proteins are provided that include (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508 wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; and (b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508, wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; wherein the first polypeptide and the second polypeptides bind to different targets; wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

Description

[0001] UW 50204.02W02 Minibinder Heterofusion Proteins induce Transdifferentiation

[0002] Federal Funding Statement

[0003] This invention was made with government support under Grant No. W81XWH-21-1-0006, awarded by the Department of Defense. The government has certain rights in the invention.

[0004] Sequence Listing Statement

[0005] The instant application contains an electronic Sequence Listing that has been submitted electronically and is hereby incorporated by reference in its entirety. The Sequence Listing was created on March 24, 2026, is named “24-2309-WO_ST26.xml” and is 1,436,588 bytes in size.

[0006] Background

[0007] The direct conversion, or transdifferentiation, of somatic non-muscle cells into skeletal myocytes offers substantial therapeutic potential for treating conditions such as muscle atrophy and sarcopenia. Cell fate transitions are often orchestrated by growth factors that induce the assembly of specific receptor pairs, leading to the transactivation of intracellular kinase domains and the initiation of lineage-specific signaling cascades.

[0008] However, natural growth factors have not been shown to induce fibroblast-to-myoblast conversion, suggesting that new strategies are needed to unlock alternative cell identities.

[0009] Summary

[0010] In a first aspect, the disclosure provides fusion proteins, comprising:

[0011] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508. wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; and

[0012] (b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508, wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; wherein the first polypeptide and the second polypeptides bind to different targets;

[0013] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0014] In one embodiment, the fusion protein comprises

[0015] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the first polypeptide binds to Human Epidermal Growth Factor Receptor 2 (Her2); and

[0016] (b) a second polypeptide comprising:

[0017] (i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 or 410-1031, wherein the second polypeptide binds to Fibroblast Growth Factor Receptor (FGFR); or

[0018] (b) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 39, wherein the second polypeptide binds to Transforming Growth Factor Beta Receptor 2 (TGFBR2);

[0019] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0020] In some embodiments, the amino acid linker is present between the first and second polypeptides. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In a further embodiment, the GS-rich amino acid linker comprises (GSGSGSGSGS)n, wherein n = 1-7 (SEQ ID NO: 1509). In another embodiment, the fusion protein comprises the amino acid sequence selected from SEQ ID NOS: 12-15 and 1457.

[0021] In another embodiment, the disclosure provides fusion proteins, comprising:

[0022] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, wherein the first polypeptide binds to Tropomyosin receptor kinase A (TrkA); and

[0023] (b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acidsequence of SEQ ID N0S:31 or 32, wherein the second polypeptide binds to Bone Morphogenetic Protein Receptor Type 2 (BMPR2);

[0024] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0025] In one embodiment, the first polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NOS:31 or 32. In another embodiment, the first polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises the amino acid sequence of SEQ ID NOS:31 or 32. In a further embodiment, the first polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16, and the second polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:31.

[0026] In some embodiments, the amino acid linker is present between the first and second polypeptides. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In a further embodiment, the GS-rich amino acid linker comprises (GGSGGSGGSG)n (SEQ ID NO: 1478), wherein n = 1-7. In another embodiment, the fusion protein comprises the amino acid sequence selected from SEQ ID NOS:33-36.

[0027] In another embodiment, the fusion proteins comprise:

[0028] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38, wherein the first polypeptide binds to Activin receptor-like kinase 1 (Alkl); and

[0029] (b) a second polypeptide comprising

[0030] (i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the second polypeptide binds to HER2; or (ii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357:, wherein the second polypeptide binds to Epidermal Growth Factor Receptor (EGFR); or(iii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359, wherein the second polypeptide binds to interleukin-2 receptor subunit gamma (IL-2RG); or

[0031] (iv) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1360-1448, wherein the second polypeptide binds to IL-2 receptor βγc heterodimer; or

[0032] (v) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408, wherein the second polypeptide binds to insulin receptor; or

[0033] (vi) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508, wherein the second polypeptide binds to Insulin-like Growth Factor Receptor 2 (IGFR2);

[0034] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.

[0035] In various further embodiments, the disclosure provides fusions proteins comprising: (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence SEQ ID NO:39that bind to Transforming Growth Factor Beta Receptor 2 (TGFBR2); or

[0036] (b) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to GammaC and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to Platelet-Derived Growth Factor Receptor (PDGFR); or(c) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to interleukin-2 receptor subunit gamma (IL-2RG) and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2; or

[0037] (d) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2 and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to PDGFR; or

[0038] (e) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408that bind to insulin receptor, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG;

[0039] (f) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1032-1035that bind to Gpl30;

[0040] (g) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:16-30that bind to TrkA, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2;

[0041] (h) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30that bind to TrkA;

[0042] (i) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2;

[0043] (j) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39 that bind to TGFBR;

[0044] (k) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2 and SEQ ID NOS: 11 and 410-103 Ithat bind to FGFR;

[0045] (l) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:1360-1448that bind to IL-2 receptor βγc heterodimer, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR; and

[0046] (m) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508thatbind to IGFR2;

[0047] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.In some embodiments, the amino acid linker is present. The linker may be any amnio acid linker suitable for an intended use. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In another embodiment, the GS-rich amino acid linker comprises (GSGSGSGSGS)n, wherein n = 1-7.

[0048] In further embodiments, the fusion protein comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500, wherein optional residues may be present or may be deleted in whole or in part. In another embodiment, the fusion protein comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500.

[0049] In another aspect the disclosure provides nucleic acids encoding the fusion protein of any embodiment or combination of embodiments of the disclosure. In a further aspect, the disclosure provides expression vectors comprising the nucleic acid of any aspect of the disclosure operatively linked to a suitable control sequence, such as a promoter. In another aspect, the disclosure provides host cells that comprise the fusion protein nucleic acid or expression vector (i.e.: episomal or chromosomally integrated) disclosed herein, wherein the host cells can be either prokaryotic or eukaryotic.

[0050] In another aspect, the disclosure provides compositions, comprising at least a first and a second polypeptide according to any embodiment or combinations of embodiments herein (i.e., Tables 1-13D), wherein the first and second polypeptides bind to different targets, and wherein the first and second polypeptides are not covalently linked.

[0051] In various embodiments:

[0052] (a) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 and 410-1031 that bind to FGFR, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl; or

[0053] (b) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to theamino acid sequence selected from SEQ ID NOS: 16-30 that bind to TrkA, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl.

[0054] In another embodiment, the (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11 or 1476;, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:37.

[0055] In another embodiment, the disclosure provides pharmaceutical compositions, comprising:

[0056] (a) the fusion protein, nucleic acid, expression vector, host cell, and / or composition of any embodiment or combination of embodiments herein; and

[0057] (b) a pharmaceutically acceptable carrier.

[0058] In another aspect, the disclosure provides methods for conversion of fibroblasts to the myogenic fate, comprising contacting fibroblasts with an amount effective of the fusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to convert the of fibroblasts to the myogenic fate. In a further aspect, the disclosure provides methods treating or limiting development of a disorder selected from muscle atrophy post-injury and muscle degeneration disorders (e.g. muscular dystrophy and sarcopenia), comprising administering to a subject in need thereof an amount effective of the fusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to treat or limit development of the disorder.

[0059] Description of the Figures

[0060] Figure 1. De novo design of novokines. (A) Previously designed minibinders against FGFR2 (PDB ID: 7TYD), TrkA (PDB ID: 7N3T), EGFR (PDB ID: 3NJP) and IGF2R (PDBID: 6UM2). (B) Flexibly linked novokines. (C) HER2 receptor with Affibody (PDB ID: 3MZW) superimposed together with HER2 minibinder. Zoom ins show key residues important for minibinder-HER2 receptor interaction. Right: Comparison of binding between the affibody (top) and the minibinder (bottom) to the HER2 receptor using BLI measurements.

[0061] Figure 2. Designed novokines enhance skeletal muscle reprogramming. (A) Schematic showing human foreskin fibroblast (HFF-iMYOD) harvested on d7 and dl4 of continuous doxycycline treatment for single cell sequencing. (B) percentage distribution of cells in full myogenic, partial myogenic or exit state post d7 or dl4 of MYOD inducible expression. (C) UMAP showing pseudotime trajectory analysis of fibroblast to commitment, partial or exit state post d7 or dl4 of MYOD inducible expression. (D) Schematic showing novokine screen method in HFF-iMYOD derived myogenic transdifferentiation. Volcano plot analyzing differential expressed genes in tSKM vs HFF-iMYOD cells in bulk RNA sequencing analysis. (E) Heat map of desmin intensity on immunostaining of tSKMs in novokine treatment in comparison to control (n=2), in two independent primary screens of designed novokines in myogenic transdifferentiation. (F) Heatmap showing various phosphoeffectors analyzed in flow cytometry of EA.hy926 cells upon novokine treatment for 15 min. (G) Confocal image showing desmin staining, MHC, and DAPI in control and HER2mb: FGFRmb (H2F) treated conditions in the myogenic transdifferentiation assay (D). scale bar=30 um (H) Schematic showing of screening method for novokines in human iPSCs-derived myogenic differentiation. (I) Desmin Intensity heat map of iSKMs treated with novokines in comparison to WTC-iMyoD. (J) In various lOOnM Novokine treatments that are positive for showing elongated myotube formation are represented. Myotube length has been significantly increased in comparison to no minibinder treatment (measured in micron). (K) Confocal images showing desmin staining, MHC, and DAPI in control, H2F treated conditions in the iPSC derived myogenic differentiation assay, scale bar=50 um. ****p < 0.0001. (L) Heatmap showing myotube desmin intensity compared to the control in two independent secondary screens of designed novokines in the myogenic trans-differentiation assay. Select novokines were screened for activity in flow cytometry phospho-effector screens.

[0062] Figure 3. HER2 and FGFRl / 2c heterodimerization shows enhanced myogenic reprogramming. (A) AlphaFold-generated predicted structures of the HER2 receptor minibinder fused with a flexible linker to FGFRlc minibinder in the orientation HER2mb: FGFRcmb (H2F) when bound to the extracellular domains of their cognatereceptors. (B) Schematic showing screen method for several HER2-RTKs novokines in human foreskin fibroblast (HFF-iMYOD) derived myogenic transdifferentiation. (C) Heatmap showing various natural ligands and HER2aff RTK novokines, do not increase desmin intensity relative to control whereas H2F, and FGFRmb: HER2mb (FH2mb) shows increased desmin intensity. (D) Confocal images showing desmin staining, MHC, and DAPI in control, H2F, and FH2mb treated tSKM. scale bar=5 um. (E) Graph showing that H2F treatment increases the conversion efficiency (percentage of nuclei in desmin positive cells) of the myogenic transdifferentiation assay. (F) Graph of the percentage of myotubes with one, two, or three and greater number of nuclei. Shows H2F and H2affF treatment significantly increases multinucleation. (G) Graph showing that H2F / FH2mb / H2affF treated tSKM show increased length compared to control (H) A volcano map showing mRNA expression of various muscle markers are upregulated in tSKM treated with H2F. (I) Schematic showing method for the long-term maturation (30 days) assay of H2F tSKM. (J) Confocal Images of H2F long term maturation tSKM stained for DAPI and alpha-actinin, scale bar=5 um. H2F treatment led to significant increase in the maturation of the myotubes as alpha-actinin striation was significantly increased in H2F treated tSKMs. Graph showing percentage of myotubes with alpha-actinin positive sarcomeres. ***p < 0.001.

[0063] Figure 4. H2F bifurcates RTK mediated MAPK and PLCy / Ca signaling response. (A) FRET performed in CHO cells expressing human HER2AC-mEYFP and FGFR1 AC-mCherry™ A scatter plot showing FRET efficiency (Eapp) significantly increased in presence of 500nM H2F in CHO cells expressing human HER2-mEYFP (donor) and FGFRlc-mCherry (acceptor) compared to PBS control. (B) Schematic showing Her2 and FGFR1 homo / heterodimerization in presence of the various H2F, FF, H2H2, FGF2 or competition with Fmb / H2mb.(C) Schematic showing phosphoproteomics flow. (D) MAPK signaling cascade analysis analyzed from phosphoproteomics data in 10 min of H2F, FGF2 and FBS treatment in CHO cells expressing human FGFRlc and Human Her2 receptors. Shows list of MAPK phosphosites shared between H2F and FGF2.(E)Graph of H2F titration curve in CHO cells expressing both human HER2_FGFRlc, HER2 FGFR1K514M, HER2K753M FGFR1. immunoblot showing pERK signal upon 50nM H2F treatment for 10 min in HER2 FGFR1K514M and HER2K753M FGFR1 expressing CHO cells. (F) Graph of H2F titration curve in CHO cells expressing HER2 FGFR1K514M,

[0064] HER2K753M FGFR1 of pAKT and Pp38. Immunoblot showing Pp38 signal upon 50nM H2F treatment for 10 min in HER2 FGFR1K514M and HER2K753M FGFR1 expressing CHO cells. (G) Model showing selective kinase domain small molecule inhibitors, Tucatiniband PD173074 against HER2 and FGFR respectively. Western blot depicting the effect of PD173074 and Tucatinib treatment on H2F treatment on CHO-hHER2-hFGFR cells FGFR receptor phosphorylation. (H) pERK signal in 100nM H2affF treatment in CHO cells expressing both hHER2 and hFGFRl and either hHER2 or hFGFR. (I) H2F shows pAKT signals in CHO-hHER2-hFGFRlc cell lines but not in either hHER2 or hFGFR. (J) H2F shows pERK in CHO-hHER2-hFGFRlc cell lines. H2F in competition with either the HER2mb or FGFRlcmb (monomeric minibinders used in excess to block one half of H2F) shows reduced pERK. (K) Heatmap showing fold change in pAKT signal in FACS sorted cells compared to PBS in EA.hy926 cells; competition with either of the Hmb or Fmb reduced the pAKT signal. Homodimers such as H2mb: H2mb or Fmb: Fmb do not show strong pAKT signals. (L) Representative images of timelapse showing Ca2+ signaling using Calbryte dye in FGF+Heparin and H2F+heparin treatments. (M) Titration curve of Ca2+ signaling using Calbryte dye in FGF+Heparin and H2F+heparin treatments. (N) Quantification of the Ca2+ signal intensity shows that FGF+heparin treatment is able to trigger calcium release in CHO-hHER2-hFGFRlc cells, whereas treatment with H2F+heparin is not. (O) Model showing that HER2mb: FGFRlcmb phosphorylates the Y653 / 654 residues of FGFR, activating the MAPK / pAKT pathways, but fails to show Ca signaling response. Thus, H2F bifurcates RTKs mediated MAPK and PLCy / Ca signaling response, scale bar= 50um. ***p < 0.001, ****p < 0.0001.

[0065] Figure 5. Effects of H2F on endothelial differentiation, pluripotency maintenance in iPSCs and primary myoblast reprogramming. (A) Schematic of the differentiation protocol from iPSCs to endothelial and perivascular lineages, highlighting intermediates. (B) Proportion of endothelial or perivascular cells generated at day 14 following treatment with FGF2, mb7, H2F. Error bars represent 2 independent biological repeats. Representative scatterplots of endothelial (VE-cadherin) and perivascular (PDGFR-B) markers, assayed using flow cytometry. (C)Immunofluorescence images showing PDGFR-B and VE-CAD expression at day 14 in cultures treated with FGF, H2F, or H2affiF. Scale bar: 100um. (D) Schematic of experimental setup to assess pluripotency marker expression in iPSCs after 48-hour treatment with FGF or H2F. Representative histograms of TRA-1-60 expression in iPSCs following treatment with varying concentrations of H2F, assayed using flow cytometry. Error represents 2 independent biological repeats. (E) Immunofluorescence images showing Oct4 and Nanog expression in iPSCs treated with FGF or H2F for 48 hours. Scale bar: 100um. (F) Oxygen consumption rate (OCR) across time for iPSCs treated with FGF or H2F. Timepoints indicate the addition of mitochondrial inhibitors (oligomycin;FCCP; antimycin A / rotenone). Oxygen consumption rate graph of 48 hours H2F and FGF treated iPSCs in E8 base media. Bar graphs show maximal respiration, spare respiratory capacity, and ATP Production. Graphs show no significant difference between H2F and FGF. (G) A schematic showing iPSCs treated with FGF / H2F in minimal media for 48h followed by Ectoderm / endoderm differentiation and RNA isolation. (H) H2F treated and FGF treated iPSCs undergo ectoderm and endoderm differentiation efficiently shown by a positive correlation of ectoderm markers (left graph) and endoderm marker expression (right graph) in respective differentiations. (I) Model summarizing the effects of H2F on stem cell fate:

[0066] Biased FGFRc activation maintains pluripotency, while its inhibition drives differentiation. (J) Schematic showing patient derived myoblast differentiation to myotubes. (K) Fusion efficiency of primary myoblasts in untreated and various concentrations of H2F. (L) Area coverage of primary myoblasts in untreated and various concentrations of H2F. (M) Alphaactinin intensity quantification in PBS,10nM FGF2 treated versus lOOnM H2F is compared at day 4. (N) Fusion efficiency of primary myoblasts in PBS, 10nM FGF versus 100nM H2F at day 4. (O) Immunofluorescence images showing untreated and 100nM H2F treated primary myotubes at day 6, stained for a-actinin and DAPI, scale bar= 20um. A small triangle marks the length measured in the plot profile. The graph shows the sarcomere distribution along the myofibrils in PBS versus H2F.(P) Distance between z-discs is measured in PBS versus 100nM H2F treated myofibers at day 6. N=4, PBS n=230, H2F n=453. (Q) Quantification of Z-disk signal clarity, measured as the FFT Peak Amplitude. (R) A model showing patient derived primary myoblast can be reprogrammed efficiently into muscle cells with potential of muscle atrophies treatment. ****p < 0.0001, * p < 0.05.

[0067] Figure 6. Direct reprogramming of human foreskin fibroblast into skeletal muscles. (A) tet-on inducible MyoD Human Foreskin Fibroblast treated with Doxycycline for day 7 and dayl4 to reprogram into muscle cells, where single cell RNA sequencing was performed. (B) UMAP showing single cell RNA clustering. Pseudotime analysis using Monocle™. (C) dot plot showing specific markers to each cluster. (D) cluster 2 versus cluster 3 comparison of barriers for the transition to muscle. (E) D8 (D4>8) reprogrammed tSKM shows muscle markers staining for Desmin, Myosin heavy chain, DAPI. (F) Confocal images of uninduced fibroblast and day 8 tSKM showing mitochondrial marker ATP-b synthase, mito-Orange uptake, Desmin and DAPI. (G) A graph showing ATP-b synthase intensity in fibroblast and day 8 tSKM. Normalised OCR measure in uninduced fibroblast, day4 and day8tSKM. Maximum respiration measured in uninduced fibroblast, day4 and day8tSKM. (H) tet-on inducible MyoD HFF treated with Doxycycline for day4 and treated with designedprotein for next 4 days to reprogram into muscle cells and stained for muscle markers. (I) A heatmap showing desmin intensity in two independent primary screens of designed proteins at 100nM concentration in iMYODHFF.

[0068] Figure 7. (A) AlklRmb-IGF2R_Endotag4 fusion directs the Alkl receptor together with IGF2R endosomal internalization, degraded through lysosomal trafficking. (B) CHO cells overexpressing the human AlklR are treated with the AlklRmb-IGF2R_Endotag4mb or IGF2R_Endotag4mb-AlklRmb for 24hrs. and washed out for another 24 hrs., cells are analyzed for AlklR expression by western blot analysis. The bar graph shows significant recovery of AlklR after recovery. (C) Immunofluorescence imaging shows AlklR, phalloidin and DAPI staining in the CHO cells expressing, bar graph shows the AlklR-intesity. (D) Immunofluorescence imaging shows reprogrammed tSKM under no mb (control) and Alkl-IGF2R_Endotag4 treatment shown in the schematic stained for muscle markers staining for Desmin, Myosin heavy chain, DAPI in PBS and Alkl. A bar graph showing fibroblast to muscle conversion efficiency is significantly upregulated in Alkl-IGF2R_Endotag4 treatment.

[0069] Figure 8. Immunofluorescence images showing No mb, C2 DPC, and C6 DPC treated 8 days old tSKM stained for Desmin, MHC and DAPI. A bar graph showing significantly increased conversion efficiency in C2 DPC and C6 DPC treated fibroblast (B) Length of myotubes significantly increased in C2 DPC and C6 DPC treated tSKM. (C) Width of myotubes significantly increased in C2 DPC and C6 DPC treated tSKM. (D) Width of myotubes significantly increased in C2 DPC and C6 DPC treated tSKM. (E) A volcano map showing genes upregulated or downregulated in D8 tSKM in C6 DPC treated tSKM versus no mb tSKM. Bulk RNA expression (F) A heatmap showing upregulation of top 50 transcription factors motifs enriched in the promoter of upregulated genes.

[0070] Figure 9. (A) Schematic showing Bulk RNA sequencing of tSKM and the mononucleated uncoverted cells. (B) Heatmap showing muscle, metabolic, Cell cycle gene, fibroblast and inflammation markers expression of transcript level in no mb (control), C2 DPC, C6 DPC treatments versus fibroblast during the D8 conversion. (C) Confocal images of uninduced fibroblast, day8 no mb tSKM, day8 DPC treated tSKM showing mitochondrial marker ATP-B synthase, mito-Orange uptake, Desmin, and DAPI. (D) A graph showing ATP-b synthase intensity in uninduced fibroblast, day8 no mb tSKM, day8 DPC treated tSKM. (E) Sarcomere formation in day30 no mb tSKM and DPC treated tSKM. (F) Single cell RNA sequencing of original resting fibroblast, no mb and C6 DPC treated mononucleated cells. UMAP showing all the nuclei sequenced. UMAP showing thefibroblast (left) and, no mb and C6 DPC (combined, right) inflammation score based on inflammatory cytokines and receptor expression level. (G) Schematic showing HFF-iMYOD induced with doxycycline for 4 days, followed by TNF-alpha treatment for next 4 days to induce inflammation followed by treatment of C6 DPC for another 4 days.

[0071] Immunofluorescence images show the shows reprogrammed tSKM various treatments shown in the schematic stained for muscle markers staining for Desmin, Myosin heavy chain, DAPI. A bar graph showing fibroblast to muscle conversion efficiency in various treatments. (H) A schematic model showing the conversion of fibroblast to skeletal muscles requires low inflammation.

[0072] Figure 10. C6 DPC treatment enhances iPSCs differentiation into skeletal muscles and enhances contractile force and accelerates kinetics in iPSC-derived EMTs.

[0073] (A) Schematic showing iPSCs-iMYOD differentiation into myotubes in C6 based designed protein cocktail for 4 days and fixed and stained for muscle makers. (B) iPSCs differentiated into myotubes are stained for DAPI, desmin and MHC. Scale bar= 10um. Violin plot showing average myotube area coverage enhanced in DPC treatment versus no minbinder treatment. (D) Mean intensity of MHC shows increase in DPC treatment versus no minbinder treatment. (E) Schematic showing mantarray assay measuring the contractile forces where the 3D muscle tissue (EMTs) is scaffolded around a rigid and a flexible post. The contractile force generated by the tissue is measured with the displacement of a magnet in the flexible post. (F) Representative immunofluorescence cross-sections of WT and DMD engineered muscle tissues (EMTs) treated with PBS (vehicle) or C6 (100 nM). Sections are stained for myosin heavy chain, dystrophin, and nuclei. Scale bar: 200 pm. Insets show higher magnification of myotube architecture (Scale bar: 20 pm) (G) Quantification of average myotube diameter. C6 induces hypertrophy in WT but not DMD myotubes. (H) Quantification of Effective Cross-Sectional Area (CSA), representing the total area occupied by myosin-positive myotubes (I) Representative tetanic (100Hz) force traces (left) and quantification of absolute active tetanic force (right). (J) Representative twitch (0.2Hz) force traces (left) and quantification of absolute active twitch force (right). (K) Specific force calculations normalizing active force to effective CSA for twitch (left) and tetanic (right) stimulation (L) Normalized relaxation traces (left) and quantification of the time to 80% relaxation (RT80%) (right), indicating improved relaxation kinetics with C6 treatment. Data are presented as mean ± S. E. M. *P < 0.05, **P < 0.01, ***p < 0.001, ****p < 0.0001 by two-way ANOVA with Tukey’s multiple comparisons test.Figure 11. A schematic showing enhanced muscle transdifferntiation, iPSC derived muscle differentiation and increased contractile forces in the 3D EMTs formed from the DMD-IPSC derived myoblast in presence of the C6 DPC.

[0074] Figure 12. Mechanism of Novokines with Alklmb and TGFBR2 mb. (A)A model showing two unrelated receptors come proximity together in presence of novokine to induce nonnative signaling. (B) Class 1 of novokines that consist of ALKlmb and class 2 with TGFBR2 mb. (C)A western showing inhibition of psmadl / 5 signaling by ALKlmb-consisting of novokines. (D) A western showing inhibition of psmad2 signaling by TGFBR2 mb consisting of novokines. (E) A western showing show degradation of ALK1 receptor in CHO cells-overexpressing human ALK1 receptor treated with Lytac-ALKl (IGF2-Endotag_4-Alkl) novokine for 24 hrs. (F) A western showing degradation of ALK1 receptor in CHO cells-overexpressing human ALK1 receptor treated with ALKl-lytac (Alkl- IGF2-Endotag_4) novokine for 24 hrs. Then recovery of ALK1 takes place in next 24 hours of washing of the novokine. (G) Immunofluorescence images of CHO cells-human ALK1R stained for ALK1R, DAPI, and Phalloidin. Images show degradation of ALK1 receptor in CHO cells-overexpressing human ALK1 receptor treated with ALKl-lytac (Alkl- IGF2-Endotag_4) novokine for 24 hrs. Then recovery of ALK1 takes place in next 24 hours of washing of the novokine. (H) A western showing show degradation of ALK1 receptor in CHO cells-overexpressing human ALK1 receptor treated with Lytac-ALKl (IGF2-Endotag_4- Alkl) novokine for 24 hrs. Then recovery of ALK1 takes place in next 24 hours of washing of the novokine.

[0075] Figure 13. TrkAmb: BMPR2mb shows improved myogenic conversion. (A) Alpha fold predicted structure of TAB2 heterofusion. (B) Image showing multinucleated myotubes on TAB2 treatment. (C) Graph showing conversion efficiency of fibroblast into myotubes. (D) Percentage of 1,2 >3 nucleated myotubes in control and TAB2. (E) Myotube length increases in TAB2 treatment. (F) Mito-Orange intensity goes up in TAB2. (G) Titration of TAB2 in CHO cells expressing human TrKA and human BMPR2. (H) Western showing CH0-TRKA-BMPR2 signals for pERK dramatically. (I)TRKA kinase inhibitor blocks TAB2 pERK activity. (J) In EA.hy926 cells pCREB signals require both receptors.

[0076] Detailed Description

[0077] All references cited are herein incorporated by reference in their entirety. Within this application, unless otherwise stated, the techniques utilized may be found in any of several well-known references such as: Molecular Cloning: A Laboratory Manual (Sambrook, et al.,1989, Cold Spring Harbor Laboratory Press), Gene Expression Technology (Methods in Enzymology, Vol. 185, edited by D. Goeddel, 1991. Academic Press, San Diego, CA), “Guide to Protein Purification” \n Methods in Enzymology (M. P. Deutshcer, ed., (1990) Academic Press, Inc.); PCR Protocols: A Guide to Methods and Applications (Innis, et al. 1990. Academic Press, San Diego, CA), Culture of Animal Cells: A Manual of Basic Technique, 2ndEd. (R. I. Freshney. 1987. Liss, Inc. New York, NY), Gene Transfer and Expression Protocols, pp. 109-128, ed. E. J. Murray, The Humana Press Inc., Clifton, N. J.), RosettaCommons.org, and the Ambion 1998 Catalog (Ambion, Austin, TX).

[0078] As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0079] As used herein, the amino acid residues are abbreviated as follows: alanine (Ala; A), asparagine (Asn; N), aspartic acid (Asp; D), arginine (Arg; R), cysteine (Cys; C), glutamic acid (Glu; E), glutamine (Gln; Q), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

[0080] In all embodiments of polypeptides disclosed herein, any N-terminal methionine residues are optional (i.e.: the N-terminal methionine residue may be present or may be deleted).

[0081] All embodiments of any aspect of the disclosure can be used in combination, unless the context clearly dictates otherwise.

[0082] Unless the context clearly requires otherwise, throughout the description and the claims, the words ‘comprise’, ‘comprising’, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. Words using the singular or plural number also include the plural and singular number, respectively. Additionally, the words “herein,” “above,” and “below” and words of similar import, when used in this application, shall refer to this application as a whole and not to any particular portions of the application.

[0083] In a first aspect, the disclosure provides fusion proteins, comprising:

[0084] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410- 1448, and 1502-1508. wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; and(b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508, wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; wherein the first polypeptide and the second polypeptides bind to different targets;

[0085] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0086] The fusion proteins of this aspect are shown in the examples to, for example, direct cell reprogramming by inducing receptor proximity, resulting in conversion of fibroblasts to the myogenic fate, and can thus be used, for example, to treat or limit development of muscle atrophies post injury, muscle degeneration disorders (including but not limited to muscular dystrophy and sarcopenia), and to develop bioengineered muscle tissues for transplantation and disease modeling. Details can be found in the examples that follow.

[0087] The amino acid sequences of SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508 are provided in Tables 1-14 below. In summary:

[0088] Table 1 provides the amino acid sequences of polypeptides that bind to Human Epidermal Growth Factor Receptor 2 (Her2) (SEQ ID NOS: 1-10). See WO / 2026 / 006160 (incorporated by reference herein in its entirety) for details on the Her2 binding polypeptides in Table 1.

[0089] Table 2 provides the amino acid sequences of polypeptides that bind to Activin receptor-like kinase 1 (Alkl) (SEQ ID NOS:37-38). See PCT / US2026 / 015090.

[0090] Table 3 provides the amino acid sequence of a polypeptide that binds to Transforming Growth Factor Beta Receptor 2 (TGFBR2) (SEQ ID NO:39). See WO / 2024 / 191776 (incorporated by reference herein in its entirety) for details on TGFBR2 binding.

[0091] Tables 4 and 4A provide the amino acid sequences of polypeptides that bind to Insulin-like Growth Factor Receptor (IGFR) (SEQ ID NOS:40-51, 43-125, and 1508). See WO / 2025 / 038596 and WO / 2023 / 288191 (each incorporated by reference herein in their entirety) for details on IGFR binding.

[0092] Tables 5A-C provide the amino acid sequences of polypeptides that bind to insulin receptor (SEQ ID NOS: 99-135, 137-154, and 156-408). See WO / 2023 / 288191 (incorporated by reference herein in its entirety) for details on insulin receptor binding.Table 6 provides the amino acid sequences of polypeptides that bind to Bone Morphogenetic Protein Receptor Type 2 (BMPR2) (SEQ ID NOS:31-32). See PCT / US2026 / 015090.

[0093] Tables 7 and 7A provide the amino acid sequences of polypeptides that bind to Fibroblast Growth Factor Receptor (FGFR) (SEQ ID NOS: 11 and 410-1031). See WO / 2023 / 288191 (incorporated by reference herein in its entirety) for details on FGFR binding.

[0094] Table 8 provides the amino acid sequences of polypeptides that bind to Glycoprotein 130 (GP130) (SEQ ID NOS: 1032-1035). See WO / 2025 / 101453 (incorporated by reference herein in its entirety) for details on GP130 binding.

[0095] Tables 9 provides the amino acid sequences of polypeptides that bind to Tropomyosin receptor kinase A (TrkA) (SEQ ID NOS: 16-30). See WO / 2023 / 288191 (incorporated by reference herein in its entirety) for details on TrkA binding.

[0096] Table 10 provides the amino acid sequences of polypeptides that bind to Platelet-Derived Growth Factor Receptor (PDGFR) (SEQ ID NOS: 1036-1335). See WO / 2023 / 288191 (incorporated by reference herein in its entirety) for details on PDGFR binding.

[0097] Table 11 provides the amino acid sequences of polypeptides that bind to Epidermal Growth Factor Receptor (EGFR) (SEQ ID NOS: 1336-1357). See WO / 2023 / 288191 (incorporated by reference herein in its entirety) for details on EGFR binding.

[0098] Table 12 provides the amino acid sequences of polypeptides that bind to interleukin-2 receptor subunit gamma (IL-2RG) (also referred to herein as Gamma C) (SEQ ID NOS: 1358-1359). See PCT / US2025 / 061404.

[0099] Tables 13-13D provide the amino acid sequences of polypeptides that bind to IL-2 receptor βγc heterodimer (SEQ ID NOS: 1360-1448). See WO / 2020 / 005819 (incorporated by reference herein in its entirety) for details on IL-2 receptor βγc heterodimer binding.

[0100] Table 14 provides amino acid sequences that bind to other targets.

[0101] For example, the first polypeptide in the fusion protein may comprise an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, the Table 1 binders that bind to Her2, and the second polypeptide in the fusion protein may comprise an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 and 410-1031, the Table 7 and 7A binders that bind to FGFR. Those of skill in the art will clearly understand the other combinations that may be made.

[0102] In some embodiments, the first polypeptide is N-terminal to the second polypeptide in the fusion protein. In other embodiments, the first polypeptide is C-terminal to the second polypeptide in the fusion protein.

[0103] In one embodiment, the fusion protein comprises

[0104] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the first polypeptide binds to Human Epidermal Growth Factor Receptor 2 (Her2); and

[0105] (b) a second polypeptide comprising:

[0106] (i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 or 410-1031, wherein the second polypeptide binds to Fibroblast Growth Factor Receptor (FGFR); or

[0107] (b) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 39, wherein the second polypeptide binds to Transforming Growth Factor Beta Receptor 2 (TGFBR2);

[0108] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0109] The fusion proteins of this embodiment are shown in the examples to, for example, direct cell reprogramming by inducing HER2-FGFR proximity, resulting in conversion of fibroblasts to the myogenic fate, and can thus be used, for example, to treat or limit development of muscle atrophies post injury, muscle degeneration disorders (e.g. muscular dystrophy and sarcopenia, and to develop bioengineered muscle tissues for transplantation and disease modeling. Details of these studies can be found in the examples that follow. In one embodiment, the first polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, and the second polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from SEQ ID NOS: 11 or 410-1031. In another embodiment, the first polypeptide comprises the amino acid selected from SEQ ID NOS: 1-10, and the second polypeptide comprises the amino acid sequence selected from SEQ ID NOS: 11 or 410-1031. In a further embodiment, the first polypeptide comprises an amino acid sequence at least80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:2. In one embodiment, the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11 or 1031.

[0110] In some embodiments, the amino acid linker is present between the first and second polypeptides. The linker may be any amnio acid linker suitable for an intended use. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In a further embodiment, the GS-rich amino acid linker comprises (GSGSGSGSGS)n, wherein n = 1-7 (SEQ ID NO: 1509). In another embodiment, the fusion protein comprises the amino acid sequence selected from SEQ ID NOS: 12-15 and 1457.

[0111] FG FRmb__H e r 2 mb: DRRKEMDKVYRT AYKR I T S T P DKE KRKE WKE AT E QLRR I AKDE E E K KKAAYM I S FLKTLGGSGSGSGSGSSVDEKIEELYEKVKE LAKKGDREATAKVLAE L YRLAVQ SGDDKVFDRLDEAYQTARENLS (SEQ ID NO: 12 )

[0112] FG FRmb_H e r 2. mb: MSGDRRKEMDKVYRT AYKR I T S T P DKE KRKE WKE AT E QLRR I AKDE EEKKKAAYMISFLKTLGGSGSGSGSGSSVDEKIEELYEKVKELAKKGDREATAKVLAELYRL AVQSGDDKVFDRLDEAYQTARENLSGSGSHHWGSTHHHHHH ( SEQ ID NO: 13 )

[0113] Her2mb__FGFRmb:

[0114] SVDEKIEELYEKVKELAKKGDREATAKVLAELYRLAVQSGDDKVFDRLDEAYQTARENLSGS GSGSGSGSDRRKEMDKVYRTAYKR I T S T P DKE KRKE KE AT E QLRR I AKDE E E KKKAAYM I SFLKTLG (SEQ ID NO: 14 )

[0115] H e r 2 mb FG FRmb:

[0116] MSGSVDEKIEELYEKVKELAKKGDREATAKVLAELYRLAVQSGDDKVFDRLDEAYQTARENL SGSGSGSGSGSDRRKE DKVYRTAYKRITSTPDKEKRKEWKEATEQLRRIAKDEEEKKKAA YMI S FLKTLGGSGSHHWGSTHHHHHH (SEQ ID NO: 15 )

[0117] In another embodiment, the disclosure provides fusion proteins, comprising:

[0118] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, wherein the first polypeptide binds to Tropomyosin receptor kinase A (TrkA); and

[0119] (b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acidsequence of SEQ ID N0S:31 or 32, wherein the second polypeptide binds to Bone Morphogenetic Protein Receptor Type 2 (BMPR2);

[0120] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

[0121] The fusion proteins of this aspect can be used, for example, to direct cell reprogramming in conversion of fibroblasts to the myogenic fate. The TrkAmb sequences are disclosed in WO2023288191, incorporated by reference herein in its entirety. The BMPR2 sequences are disclosed in PCT / US2026 / 015090.

[0122] In one embodiment, the first polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NOS:31 or 32. In another embodiment, the first polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises the amino acid sequence of SEQ ID NOS:31 or 32. In a further embodiment, the first polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16, and the second polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:31.

[0123] In some embodiments, the amino acid linker is present between the first and second polypeptides. The linker may be any amnio acid linker suitable for an intended use. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In a further embodiment, the GS-rich amino acid linker comprises (GGSGGSGGSG)n (SEQ ID NO: 1478), wherein n = 1-7. In another embodiment, the fusion protein comprises the amino acid sequence selected from SEQ ID NOS:33-36.

[0124] > TrkAmb: BMPR2mb RDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIRQVEKRLRSGG SGGSGGSGTEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWRILVQLVEAAKE AEEIN (SEQ ID NO: 33)

[0125] TrkAmb: BMPR2mbMSGRDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIRQVEKRLR SGGSGGSGGSGTEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWRILVQLVEA AKE AE E I NGSGSHHWGSTHHHHHH (SEQ ID NO: 34 )

[0126] BMPR22mb-TrkAmb MSGTEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWRILVQLVEAAKEAEEIN GGSGGSGGSGRDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIR QVEKRLRS (SEQ ID NO: 35)

[0127] BMPR22mb-TrkAmb TEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWRILVQLVEAAKEAEEINGGS GGSGGSGRDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIRQVE KRLRS GSGSHHWGSTHHHHHH (SEQ ID NO: 36)

[0128] In another embodiment, the fusion proteins comprise:

[0129] (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38, wherein the first polypeptide binds to Activin receptor-like kinase 1 (Alkl); and

[0130] (b) a second polypeptide comprising

[0131] (i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the second polypeptide binds to HER2; or (ii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357:, wherein the second polypeptide binds to Epidermal Growth Factor Receptor (EGFR); or

[0132] (iii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359, wherein the second polypeptide binds to interleukin-2 receptor subunit gamma (IL-2RG); or

[0133] (iv) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1360-1448, wherein the second polypeptide binds to IL-2 receptor βγc heterodimer; or(v) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408, wherein the second polypeptide binds to insulin receptor; or

[0134] (vi) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508, wherein the second polypeptide binds to Insulin-like Growth Factor Receptor 2 (IGFR2);

[0135] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.

[0136] In various further embodiments, the disclosure provides fusions proteins comprising: (a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence SEQ ID NO:39that bind to Transforming Growth Factor Beta Receptor 2 (TGFBR2); or

[0137] (b) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to GammaC and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to Platelet-Derived Growth Factor Receptor (PDGFR); or

[0138] (c) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to interleukin-2 receptor subunit gamma (IL-2RG) and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2; or

[0139] (d) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2 and asecond polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to PDGFR; or

[0140] (e) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408that bind to insulin receptor, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG;

[0141] (f) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1032-1035that bind to Gpl30;

[0142] (g) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30that bind to TrkA, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2;

[0143] (h) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30that bind to TrkA;

[0144] (i) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2;(j) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39 that bind to TGFBR;

[0145] (k) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2 and SEQ ID NOS: 11 and 410- 1031 that bind to FGFR;

[0146] (l) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:1360-1448that bind to IL-2 receptor βγc heterodimer, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR; and

[0147] (m) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508thatbind to IGFR2;

[0148] wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.

[0149] In all embodiments, the fusion proteins can be used, for example, to direct cell reprogramming by inducing receptor proximity, resulting in conversion of fibroblasts to the myogenic fate, and can thus be used, for example, to treat or limit development of muscle atrophies post injury, muscle degeneration disorders (including but not limited to muscular dystrophy and sarcopenia), and to develop bioengineered muscle tissues for transplantation and disease modeling.

[0150] In one embodiment of all of these embodiments, the first polypeptide and the second polypeptide comprise an amino acid sequence at least 80% identical to the reference sequence. In another embodiment, the first polypeptide and the second polypeptide comprisean amino acid sequence at least 90% identical to the reference sequence. In a further embodiment, wherein the first polypeptide and the second polypeptide comprise an amino acid sequence at least 95% identical to the reference sequence.

[0151] In some embodiments, the amino acid linker is present. The linker may be any amnio acid linker suitable for an intended use. In one embodiment, the amino acid linker comprises a GS-rich amino acid linker. In another embodiment, the GS-rich amino acid linker comprises (GSGSGSGSGS)n, wherein n = 1-7.

[0152] In further embodiments, the fusion protein comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500, wherein optional residues may be present or may be deleted in whole or in part. The fusion protein sequences of tis embodiment are shown in Table 15, with residues in parentheses being optional. In another embodiment, the fusion protein comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500.

[0153] In another aspect the disclosure provides nucleic acids encoding the fusion protein of any embodiment or combination of embodiments of the disclosure. The nucleic acid sequence may comprise single stranded or double stranded RNA or DNA in genomic or cDNA form, or DNA-RNA hybrids, each of which may include chemically or biochemically modified, non-natural, or derivatized nucleotide bases. Such nucleic acid sequences may comprise additional sequences useful for promoting expression and / or purification of the encoded peptide or chimeric molecular construct, including but not limited to polyA sequences, modified Kozak sequences, and sequences encoding epitope tags, export signals, and secretory signals, nuclear localization signals, and plasma membrane localization signals. It will be apparent to those of skill in the art, based on the teachings herein, what nucleic acid sequences will encode the polypeptide or fusion protein of the disclosure.

[0154] In a further aspect, the disclosure provides expression vectors comprising the nucleic acid of any aspect of the disclosure operatively linked to a suitable control sequence, such as a promoter. “Expression vector” includes vectors that operatively link a nucleic acid coding region or gene to any control sequences capable of effecting expression of the gene product. “Control sequences” operably linked to the nucleic acid sequences of the disclosure are nucleic acid sequences capable of effecting the expression of the nucleic acid molecules. The control sequences need not be contiguous with the nucleic acid sequences, so long as they function to direct the expression thereof. Thus, for example, intervening untranslated yettranscribed sequences can be present between a promoter sequence and the nucleic acid sequences and the promoter sequence can still be considered “operably linked” to the coding sequence. Other such control sequences include, but are not limited to, polyadenylation signals, termination signals, and ribosome binding sites. Such expression vectors can be of any type, including but not limited plasmid and viral-based expression vectors. The control sequence used to drive expression of the disclosed nucleic acid sequences in a mammalian system may be constitutive (driven by any of a variety of promoters, including but not limited to, CMV, SV40, RSV, actin, EF) or inducible (driven by any of a number of inducible promoters including, but not limited to, tetracycline, ecdysone, steroid-responsive). The expression vector must be replicable in the host organisms either as an episome or by integration into host chromosomal DNA. In various embodiments, the expression vector may comprise a plasmid, viral-based vector, or any other suitable expression vector.

[0155] In another aspect, the disclosure provides host cells that comprise the fusion protein nucleic acid or expression vector (i.e.: episomal or chromosomally integrated) disclosed herein, wherein the host cells can be either prokaryotic or eukaryotic. The cells can be transiently or stably engineered to incorporate the expression vector of the disclosure, using techniques including but not limited to bacterial transformations, calcium phosphate coprecipitation, electroporation, or liposome mediated-, DEAE dextran mediated-, polycationic mediated-, or viral mediated transfection.

[0156] In another aspect, the disclosure provides compositions, comprising at least a first and a second polypeptide according to any embodiment or combinations of embodiments herein (i.e., Tables 1-13D), wherein the first and second polypeptides bind to different targets, and wherein the first and second polypeptides are not covalently linked.

[0157] As described in Example 3, combinations of the binding proteins significantly increased the efficiency of myogenic transdifferentiation in human fibroblasts.

[0158] In various embodiments:

[0159] (a) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 and 410-1031 that bind to FGFR, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl; or

[0160] (b) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30 that bind to TrkA, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl.

[0161] In another embodiment, the (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11 or 1476;, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:37.

[0162] The fusion protein, nucleic acid, expression vector, and / or host cell may be the sole active agent in the composition, or the composition may further comprise one or more other agents suitable for an intended use.

[0163] In another embodiment, the disclosure provides pharmaceutical compositions, comprising:

[0164] (a) the fusion protein, nucleic acid, expression vector, host cell, and / or composition of any embodiment or combination of embodiments herein; and

[0165] (b) a pharmaceutically acceptable carrier.

[0166] The compositions may further comprise (a) a lyoprotectant; (b) a surfactant; (c) a bulking agent; (d) a tonicity adjusting agent; (e) a stabilizer; (f) a preservative and / or (g) a buffer. In some embodiments, the buffer in the pharmaceutical composition is a Tris buffer, a histidine buffer, a phosphate buffer, a citrate buffer or an acetate buffer. The composition may also include a lyoprotectant, e.g. sucrose, sorbitol or trehalose. In certain embodiments, the composition includes a preservative e.g. benzalkonium chloride, benzethonium, chlorohexidine, phenol, m-cresol, benzyl alcohol, methylparaben, propylparaben,chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid, and various mixtures thereof. In other embodiments, the composition includes a bulking agent, like glycine. In yet other embodiments, the composition includes a surfactant e.g., polysorbate-20, polysorbate-40, polysorbate- 60, polysorbate-65, polysorbate-80 polysorbate-85, poloxamer-188, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trilaurate, sorbitan tristearate, sorbitan trioleaste, or a combination thereof. The composition may also include a tonicity adjusting agent, e.g., a compound that renders the formulation substantially isotonic or isoosmotic with human blood. Exemplary tonicity adjusting agents include sucrose, sorbitol, glycine, methionine, mannitol, dextrose, inositol, sodium chloride, arginine and arginine hydrochloride. In other embodiments, the composition additionally includes a stabilizer, e.g., a molecule which substantially prevents or reduces chemical and / or physical instability of the nanostructure, in lyophilized or liquid form. Exemplary stabilizers include sucrose, sorbitol, glycine, inositol, sodium chloride, methionine, arginine, and arginine hydrochloride.

[0167] The fusion protein, nucleic acid, expression vector, and / or host cell may be the sole active agent in the pharmaceutical composition, or the composition may further comprise one or more other agents suitable for an intended use.

[0168] In another aspect, the disclosure provides methods for conversion of fibroblasts to the myogenic fate, comprising contacting fibroblasts with an amount effective of the fusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to convert the of fibroblasts to the myogenic fate. As disclosed in the examples that follow, the fusion proteins can be used to direct cell reprogramming by inducing receptor proximity, resulting in conversion of fibroblasts to the myogenic fate, and can thus be used, for example, to treat or limit development of muscle atrophies post injury, muscle degeneration disorders (including but not limited to muscular dystrophy and sarcopenia), and to develop bioengineered muscle tissues for transplantation and disease modeling. The method can be carried out in vitro, in vivo, or ex vivo.

[0169] In a further aspect, the disclosure provides methods treating or limiting development of a disorder selected from muscle atrophy post-injury and muscle degeneration disorders (e.g. muscular dystrophy and sarcopenia), comprising administering to a subject in need thereof an amount effective of the fusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to treat or limit development of the disorder.As used herein, "treat" or "treating" a disorder means accomplishing one or more of the following in a subject with the disorder: (a) reducing the severity of the disorder; (b) limiting or preventing development of symptoms characteristic of the disorder(s) being treated; (c) inhibiting worsening of symptoms characteristic of the disorder(s) being treated; (d) limiting or preventing recurrence of the disorder(s) in patients that have previously had the disorder(s); and (e) limiting or preventing recurrence of symptoms in patients that were previously symptomatic for the disorder(s).

[0170] The subject may be any subject that has a relevant disorder or is / may be at risk of the relevant disorder. In one embodiment, the subject is a mammal, including but not limited to humans, dogs, cats, horses, cattle, etc.

[0171] As used herein, an “effective” amount refers to an amount of the fusion protein, nucleic acid, expression vector, and / or host cell that is effective for treating the disorder. The fusion proteins, nucleic acids, expression vectors, and / or host cells are typically formulated as a pharmaceutical composition, such as those disclosed above, and can be administered via any suitable route, including but not limited to orally, by inhalation spray, ocularly, intravenously, subcutaneously, intraperitoneally, and intravesicularly in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles.

[0172] Any suitable dosage range may be used as determined by attending medical personnel. Dosage regimens can be adjusted to provide the optimum desired response. A suitable dosage range for the polypeptides or fusion proteins may, for instance, be 0.1 ug / kg-100 mg / kg body weight; alternatively, it may be 0.5 ug / kg to 50 mg / kg; 1 ug / kg to 25 mg / kg, or 5 ug / kg to 10 mg / kg body weight.

[0173] The fusion proteins, nucleic acids, expression vectors, host cells, and / or pharmaceutical compositions made be administered as the sole therapeutic agent, or may be administered together with (i.e.: combined or separately) one or more other therapeutic agents,

[0174] The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While the specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize.

[0175] TablesTable 1. Herl minibinders

[0176] SEQ ID NO Name Amino acid sequence

[0177] 1 >her2 bi SVDEKIEELYEKVKELAKKGDREATAKVLAEMYRLAVASGDPKVFDRLVEAF nder par NIARENLS

[0178] ent

[0179] Top Her2 binders after combo optimization:

[0180] 2 >her2 cb SVDEKIEELYEKVKELAKKGDREATAKVLAELYRLAVQSGDDKVFDRLDEAY 0672~ QTARENLS

[0181] 3 >her2 cb SVDEKIEELYEKVKELAKKGDREATAKVLAELYRLAVERGDPKVIDRLGEAY 0638~ QIARENLS

[0182] 4 her2 cb S VDEKI EELYEKVKELAKKGDREATAKVLAEMYRLAQEKGDAKFI DRLGEAF 0781” KIARENLS

[0183] 5 her2 cb SVEEKIEELYEKVKELAKKGDREATAKVLAEMYRLAQASGDAKFI DRLGEAF 1032~ QKARENLS

[0184] Her2 binder variants for conjugation to cysteine:

[0185] 6 >her2 cb SVDEKIEELYEKVKCLAKKGDREATAKVLAELYRLAVQSGDDKVFDRLDEAY _0672~ 15 QTARENLS

[0186] C

[0187] Variants for conjugation to cysteine or lysine:

[0188] 7 >her2 cb SVDEKIEELYEKVKCLAKRGDREATARVLAELYRLAVQSGDDRVFDRLDEAY _0672~ 15 QTARENLS

[0189] C lys4

[0190] Her2 binder variants with altered binding affinity: 8 >her2 cb SVDEKIEELYEKVKCLAKKGDREATAKVLAELHRLAVQSGDDKVFDRLDEAY _0672~ 15 QTARENLS

[0191] C 33H

[0192] 9 her2 cb SVDEKIEELYEKVKCLAKKGDREATAKVLAELYRLAVQSGDDKVHDRLDEAY 0672~15C QTARENLS

[0193] 45H

[0194] 10 >her2 cb SVDEKIEELYEKVKCLAKKGDREATAKVLAELYRLAVQSGDDKVFDRLDEAH _0672~ 15 QTARENLS

[0195] C 52H

[0196] 1502 SVDNKFNKEMRNAYWEIALLPNLNNQQKRAFIRSLYDDPSQSANLLAEAKKL

[0197]

[0198] NDAQAP

[0199] Table 2. Alk1 binders

[0200] SEQ Name Short Sequence

[0201] ID NO name

[0202] 37 alkl vc Alkl_m GQEEI FRALALFSADLLNIEDVKIESKDGRVKVTVKGNSPDSEE 3 64~ 2 bl FERYLRELAERLGLEVEIERT

[0203] 38 alkl_60 Alkl_m GQEEI FRALALFSADLLNLSREVTELI IKLVLTGDPELEEELEK

[0204]

[0205] 4 b2 AVEENGDNREDAEEFLKYVKELAKRLGL

[0206] Table 3. TFGBR2 binders (residues in parentheses are optional)

[0207] 5HCS_TGFBR2_1 (G) LKELLKELNKAIASGDTETVRRILEELLELLKEAFEKGDYDLAISIASMAVK AASYIGDTETLKELLEILKKIKEKLKKEGDEAALKAVERNIKWEKVA ( SEQ (TGFbRII

[0208] ID NO: 39 )

[0209] binder)

[0210]

[0211] Table 4. IGFR binders

[0212] Sequence / Name TERRVIQVLEQILEDEDPEWEKMLEILLEILEEAGDPARKLVEEILRWRKNLEEARELVRRLS IGF- 2R: D6mb ( SEQ ID NO: 40 ) MVDARLLLLIWEAKELLERGNPEEARKVLEEAREIAERNNNEELLKELEVLLEKLE IGF-2R: Dllmbl ( SEQ ID NO: 41 ) MVEASLWLLIWDAGELVERGNPEEARKVLEEAREIAERNNREEFLKELEVLLEKLE IGF-2R: Dllmb2 ( SEQ ID NO: 43 ) MEEAQRLLEEWEFQERVDELARKYGERVKEVGRQIIQDPDPEVRRKMLDILERIYREA (G) ( SEQ ID NO: 44 )

[0213] IGF Tag3 ( Rigid2 ) MEEAQRLLLEWEAQEWADSQGEDAKRVKQVVQQILQDPDLEVQKKMLEILKRIYEEK (G) ( SEQ I D NO: 45 ) IGF Tag4 ( Rigid ) MKEAQRLLLEWEAREWAKSQGEDAKRVLEVMKQILQDPDLDVQRKMLEILKRIYEEKG ( SEQ ID NO: 4 6 ) IGF_Tag5 ( Rigid2 ) MEEAQRLLLEWEAREWAESQGEDSKRVKQVVEQILQDPDLEVQRKMLEILKRIYEEKG ( SEQ ID NO: 47 ) IGF Tag 6 ( Rigid3 ) MDEARRLLLEWECRERVESLAKTEEERKRMLQVLSQILEDPDVEVVEKMVDILCRIYREAG

[0214] ( SEQ ID NO: 48 MDEAKRLLFEWEARERIESLAKTEEEKKRMLQVLSQILEDPDVEVREKMVDILVRIYKEAG

[0215] ( SEQ ID NO: 49 ) MEEAKRLLFEWEARERVVSLAKTEEERKRMLQVLSQILEDPDVEVREKMVDILCRIYREAG

[0216] ( SEQ ID NQ: 50 ) IGF Tag8 ( Rigid6 ) MEEAKRLLFEWEARERVEALAKTEEERKRMLQVLEQILEDPDVEVQRKMVDILCRIYREAG

[0217] ( SEQ ID NO: 51 ) > IGF_Tag 9 ( Rigid? ) SVEERLEEIARKYLPPEDVEFLKLALRALIKDLNVPPERALKQLIWIARQSGDPELVRVLELALK

[0218] ( SEQ ID NO: 1508 )

[0219]

[0220] Table 4A. IGF1 R binding polypeptides

[0221] SEQ Sequence

[0222] ID NO:

[0223] 52 GSSTRNAEFIVLLAELCAKSQNDPSLQEYVKKVKKIVESLLSNGDEKSAEEVARKAL EYCADGGS

[0224] 53 STRNAEFIMLLLELCVKSKNDPQVQEYVKKVKKQVERLVGNGDEKKAEEVARKALEY CADG

[0225] 54 DKELLERWELLRQGNPEEARKLLRELLRELAELGDPLKRLVFQLLVLLAAGDPEWK EILKKLEK

[0226] 55 TAEELLEIAVKDGNPELLRAMLRDIARASALAGNLEELEKVLERLLKEVGDPI FEEA IRLVKELH

[0227] 56 GSDETCKEAERNIEFLALLAKLLDDPEHAERLLKQAEELAKKCNDPRLEKLVKKLSE EVRKYVGS

[0228] 57 SEELEKKVKEAVEEGDPVIVFLLLAELLAETGDPEVEKLERLLQEALRRDDPDLARE VLELVKKT

[0229] 58 GSDEKCKEAERNIEFLALLAKLLDDPEAAEELLKQAEELAKKCNDPRLEKLVKKLSE

[0230]

[0231] EVRKYVGSDEEVRRLLKELLESGRFSCFAWLLVSVDLIAKGIPPEEAAELVERAIKKLGDPCLE EALRELRT SKIEKAIKLAKELGDPDAVRILELVKEFLEEGDPERAKALLRFLILILRQFGDPELV KIVEEVLP DIVEEVIRKAEELGNPHVARLIEFLLALVKLAGLPPEELIKMLEHALEIEGDPI IRE LIELLRKK SEDDKKCKSVEDKARELIKKARDPLVQRALKAAIELIQKASQVSDPENKEFLCLLAK LLVESAIG DEELEKAVELARRLGVDPEALKALLRAIKEALERGDKENLEFLLLLLELIVNDPEVR KLVRKVIK DEEVRRLLKELLESGRFSPFAWLLVSVDLIAKGIPPEEAAELVERAIKKLGDPDLE EALRELRT SKVERLIELLKRLGDPNLRHLARALEEALRNNDEENLEFILLLIELVARALGDPEVL KLVREVLK SEDDKKLKSVEDKARELIKKSRDPEAQRALKAAIELIQEASQVSDPENKEFLVLLAH LIVEAAAG DDREFVAELILWLARFNPENAEHLCEAIVKILGLSPELEELCRRLVELVERGDKDKV EELLRRAK PLAEESVEIAKREGNLEEAIEFLLLLARLLGNEEAVRLLEKAKELLKEGDPDKAEEL VRRVLEII SDETAHRI IRRVLEICDKRGDEDCSQKVLRLAILARSDAHHASSEEEASRIWQDAAK KVQKLESG GSDLEDLKKEAKKLEKWVSNCRDPLLREAVSRLIEDVKKAADKGDKHNAEFAVLLAQ LFVKSCGS SDEQEEIRKKVQETLAELIKKCQKDPETAKKEVKKLRELAERLGDEELKRNVEFARL LCELLEGS SREEEAVRESLREALEEGDPRTVSLALAICRIILGDPDLCREILREIAKELGNEEW KEAEKLLP GSDWRQAEHILRFIQHIARAANDEDALKRATTLLKQLQEATKKGDEDTVKRVIEEAE KLLRRAGS SPLEEEVKKLVETAEKAAKDKNDPHAEEAVKSLDKLFKEAKKKNDEENLEFVSLLAR LVLRAVIG SEEAKKVLKFIKKAEKEARDANDPVAVELAHEALKLAEDAARRNDKEALRQAEFLAF LAAFIARS SKVERLIELLKRLGDPNLRHLARALEEALRNNDEENLEFILLLIELVCRALGDPECL KLVREVLK SEEDELRKLVEELEEKARKVIRTEEQREAVESLIDLAKKALKNGDKENVEFLALLVH LLIEAGGS PEIEEKFEHIMRLLPPEDIQLLIFLLMVASEKGTEEEVIRYFVELVEKLGDPKEALK LLEEYVRR PEIEEKFEHIMRLLPPEDIQLLIFLLMVASEKGTEEEVIRYFVECVEKLGDPKECLK LLEEYVRR GSDWRQAEHILRFIQHIARAANDEDALKRATTLLKQLQECTKKGDEDCVKRVIEEAE KLLRRAGS GSSTRNAEFIVLLAELLAKSQNDPSLQEYVKKCKKIVESLLSNGDEKSAEEVARKCL EYVADGGS SDERERLREAVEELLRLAREENDAFIAALAVLLARSALEQDKDDELSRRLLELAEKV ARELRSGS SFALQILLRSARHHGDEEWRVLERVIKLLEDGDPDSAEKLARELLEKTGDPI IRAA

[0232]

[0233] LELLKRLK84 SEELEKKVKEAVEEGDPVIVFLLLAELLAETGDPEVEKLERLLQECLRRDDPDLCRE VLELVKKT

[0234] 85 SDETAHRI IRRVLEIADKRGDEDLSQKVLRLAILARSDAHHASSEEEASRIWQDAAK KVQKLESG

[0235] 86 SCIEKAIKLAKELGDPDAVRILELVKEFLEEGDPERAKALLRFLILILRQFGDPELV KIVEECLP

[0236] 87 SPEDKEKARKILQSVLELVAHNQIDEKIASKLISEVKTLAKRVNDPQLLRNAEFVEL LVRVLYRG

[0237] 88 DEEVRRLLKELLESGRFSPFAWLLVSVDCIAKGIPPEECAELVERAIKKLGDPDLE EALRELRT

[0238] 89 GSSTRNAEFIVLLAELLAKSQNDPSLQEYVKKVKKIVESLLSNGDEKSAEEVARKAL EYVADGGS

[0239] 90 GSDLEDLKKEAKKLEKWVSNLRDPLLREAVSRLIEDVKKAADKGDKHNAEFAVLLAQ LFVKSDGS

[0240] 91 DEEVRRCLKELLESGRFSPFAWLLCSVDLIAKGIPPEEAAELVERAIKKLGDPDLE EALRELRT

[0241] 92 NFKEVIEELLRDGNCLIATALALTVLRELNTPEARELLKLLREALERRDPELLCRLL ELVLELLK

[0242] 93 SDERERLREAVEELLRLAREENDAFIAALAVLLARSCLEQDKDDELCRRLLELAEKV ARELRSGS

[0243] 94 GSDKELAKKAVKELLETALQQGDEENFEFAVLLAKLLARAVNDPDVERI IKELEKKE KDLKKSGS

[0244] 95 DEEELKELAKRCGIPPEEVEKVLRKLEEHFGNPEAAKAMLFLALAACVYDPDSPTAR VAREIIKK

[0245] 96 SEEERILKLLEEAVRRGDPELLEELIRLLSELLGDPRNALFLAALYAIAAGNPEAEK IVEELRRR

[0246] 97 S GS SKEDAEKLLEKLVEAARRNDDE TVTKLQQL I DE I CKKEQDDNVC FLALLAEVLL RSLKRGSG

[0247] 98 DAERLIEI IREIHKKNPEAARLLFLILFLLLLLKDPELRKLYELVREAVEKGDPELV

[0248]

[0249] RRLEKLVH

[0250] Table 5. InsulinR binding polypeptides (F1 domain)

[0251] SEQ Name Sequence

[0252] ID NO:

[0253] 99 m_HHH_b l_07075_00000 DEHYREWVKNLLEIARNLRDDEEVRHLLEEAERVAKEGNDPELEE 0000_0001_20_35_H_. _ LVRRFREEF

[0254] HHH_b2_02238_0001_00

[0255] 05

[0256] 100 m ferr buw 10729 000 DLEEANRVLMKYADLLEEVYKKDEEEAERLLREAEELLERLGDWS 000000~0001_45_53_H_ AVMLIRKLFEEL

[0257] . _HHH_b2_01165_0001_

[0258] 0002

[0259] 101 m ferr buw 10772 000 DVEELLEEARKATEEGDHEKVWELLEEAFRLAEEKNDPRLQLLVL 000000~0001_41_52_H_ EVAVELIEILE

[0260] • _HHH_b l_00 95 0_0001_

[0261] 0002 0001

[0262] 102 m bcov 4helix 09130 DVEERLEEMVEDLVKMLGLSEEQEEELRRYAEELLERGVDPRRVE 000000000_0001_17_28 SKVVNLALVLRD

[0263] _H_. _HHH_b2_03173_00

[0264]

[0265] 01 0002m ferr buw 04504 000 LEVRLENASEDLKWAAKMLAEEAGAEVEESNGRLTIEGLSEEAAL 000000~0001_43_53_H_ FLEMLAKEYGVKVEISE

[0266] . ferr buw 20197 000

[0267] 1“ 0002“ 0001

[0268] m ferr buw 06905 000 RRIEVKLSDETELWFLELFLRKNGIKVERRGDTVHIEGI SEELIE 000004~0001_10_l 9_H_ DVKFLAKVYNGRVREVK

[0269] . ferr buw 18253 000

[0270] l“0001“

[0271] HHH_bl_00008_0001_00 SEEELERLEREIRRLLKEGDLEEAHHLIFRLAELAQELNDLEAFE 04 30 IVFELFEILDKLR

[0272] m ferr buw 15156 000 SEELEELAKEIVERLVKELGLSEEQKEDLEWVAEMLLEMGESPES 000000~0001_14_22_H_ IEQFLRRVAEMLG

[0273] • _HHH_b2_07 64 1_0001_

[0274] 0001

[0275] m bcov 4helix 11996 STVTFTVDDEDEARLLADVLREVDGLRVELNGNKVTVELDNERDA 000000000_0001_40_48 HFWAHTFERTFGIPVRIRD

[0276] H. ferr buw 08589

[0277] 0001“

[0278] m ferr buw 05569 000 TTIRITTDDEEALKQLEDLAKFLGIRVELSGDKVTFHVTDEQQEF 000000~0001_10_21_H_ FLRLLIKFLGLEYRTEK

[0279] . ferr buw 01112 000

[0280] l~0002~

[0281] m_HHH_b2_04391_00000DESDE IYNEAVREAMELYDRGDPEEAERVLREAIELLRKLGNPFL 0000_0001_40_51_H_. _ AKLLELLIEYL

[0282] HHH_bl_00580_0001_00

[0283] 01

[0284] m_HHH_b2_04391_00000DESDRIYNEAVRKAMELYDRGDPEEAERVLRKAIELLRKLGNPFL 0000_0001_40_51_H_. _ AKLLELLIEYL

[0285] HHH bl 00580 0001

[0286] S2B1 SKLEEIEELLKELSKTNPLAKDILWVIEVRTEDGHDPKSELVFIR QYLKTLNTPEAREILKIVAPRF_403_s2 BSELDEIKKLLEELSKTNPLAKDILWVIDVREEDGHDPESELVFIR QYLKTLKDDPEAQEVLELFN RF_404_S2B SELDEIEELLEELSKTNPLAKDILWVIRVRREDGHDPESELVFIR QYLRTLKGSSPEADEVLALF RF_405_S2B SELEEIKKLLEELSKTDPLAKDILWVIEVREEDGHDPESELVFIR QYLKTLNEPSPARKLIESYW RF_406_S2B SELEEIKKLLKELSKTDPLAKDILWVIEVREEDGHDPESELVFIR QYLKTLNTPSPARKLIEEIH RF_407_S2B SKLDEIKELLEKLSETNPLAKDILWVIEVREEDGHDPESELVFIR QYLKTLNTPEAQEVLSVINK RF_408_S2B SKLEEIKKLLKELSKTDPLAKDILWVIEVREKDGHDPESELVFIR QYLKTLNEPSPARKLIESIF RF_409_S2B SPLDELAARLEELSKTNPLAKDLLWVIREERADGFDPEEVLRFLR NYLRNQRGKSPEADAVLALF RF_410_S2B SLLDELRKKLEELSKTNPLAKDILWVIEELEKDGEDPLKVLEFIY NYLRNLRGKSPEADEVLALL RF_411_S2B SKLDELKEKLTELAKTNPLAKDILWVIEEMEADGFDPEKVLEFIY NYLKNLRGKSPEADKVLALF RF_412_S2B SDIEKLKEKLKELSETDELAKDILWVIEVREEDGHDPESELVFIR QYLKTLNKESKAAELVFSYY RF_413_S2B SDIEKLKKELKELSKEDELAKDILWVIEVREEDGHDPESELVFIR

[0287]

[0288] QYLKTLNEKSKAAELVFKYY123RF_414_S2BSPIDKLKEKLKELSKTDPLAKDILWIFEEMEKEGEDPEKVLEFVV RYLKTEKEESPAAKLIFDYR

[0289] 124RF_415_S2BSPLEKLKKELKELSKTDELAKDILWVFEEMEKEGEDPEKVLEFIL NWLKTENEKSPSAELVFEYE

[0290] 125RF_416_S2BSDIDELLEKLRELSETDPLARDILWVFEEMLAEGEDPEKVLEFVV

[0291]

[0292] RWLKTENKESPAAKLIFDYV

[0293] Table 5B. InsulinR binding polypeptides

[0294] SEQ ID Name SIB component

[0295] NO:

[0296] 126 RF405 LIB AFEAFMKLMDALFLAKDPEIKKKAEELIKKLKEADEKGDIEE LEKVVEEAEEVYEKVK

[0297] 127 RF409 LIB PRVLKKAYDAYIKLLEALYKSKDEELKKEAEKLLKELLEASE AGDLDKLLELSEKAEEIYKKVK

[0298] 128 S2-F2-S1 and other PE VKKE AFKAFMLLMDAL FLAE DPNI RKT I EEL I ERLE KADE flexible fusions NDDEEKLKEIIKKAKTIWKQVL

[0299] ( original

[0300] sequence )

[0301] 129 S2-F1-S1 SEVKKEAFKAFMDLMDALFLAKDPEIKKKAEELIKKLEEADE KNNE E E L KKV VE E AKKVY KS VK

[0302] 130RF_403_s l BPKVKKEAFEAFMKLMDALFLAKDPEIKKKAEELIKKLLEADE KGDLEKLEEVVKEANKVYEEVK

[0303] 131 RF_4O4_S1B PKVKKRAFEAFMKLMDALFLAKDPEIKKKAEELIKKLLEADE KGDLEELEEVVKEAE KVY E KVK

[0304] 132 RF_4O5_S1B KKVKKKAFEAFMKLMDAL FLAKDPE I KKKAEEL I KKLKEADE KGDIEELEKVVEEAEEVYEKVK

[0305] 133 RF_4O 6_S1B KKVKKE AFE AFMKLMDAL FLAKDPE I KKE AE KL I KE LL KADE EGDIEKLEEVVKKANEVYEKVK

[0306] 134RF_407_S 1 BKE VKKAAF E A FMKLMD AL FL AKD P E I KKKAE E L I KE LL E ADE KGDLEKLEEVVKEAE KVY E E VK

[0307] 135 RF_4O8_S1B KE VKKRAF E A FMKLMD AL FL AKD P E I KKE AE E L I KE LL E ADE KGDIEKLEEVVERAEEVYEKVL

[0308] 137RF_410_S 1 BPLVLAEGYEAYLLLLETLVESKDPELREEAERLIEELLAANE AGDLDRIREVVRRAREVYEKVK

[0309] 138 RF_411_S1B PFVLEEAYKAYLKLLETLEKSKDPELKKEAEKLIERLLAANE AGDLDALREAVREAEKLYEKVK

[0310] 139 RF_412_S1B KKVKKKAFEAFMKLMDAL FESKDEELKKKAEELIKKLKEADE KGDVELLEELVEEAEKVYKEVK

[0311] 140RF_413_S 1 BKKVKKKAFEAFMKLMDAL FKSKDEELKKKAEELIKKLKEADE KG D VE E L E KV VE E AE E VY KKVE

[0312] 141RF_414_S 1 BKKVLEEALEAYYELLRVLEESKDPELKKRAEALIEELLRANA EGDIEKLEELVREAKELAAEAK

[0313] 142RF_415_S 1 BKRVLEEAYEAYLELLEVLEKSKDPDLKKAAEELLERLRRANE EGNIEELRELVREAKELAEKAK

[0314] 143RF_41 6_S 1 BKKVLEEAYKAWYALLDVLVKSKDPALKAEAEALLAGLRAANE

[0315]

[0316] AGDVEKLRELVAEAKKVAAKAA

[0317] Table 5C. InsuUnR binding polypeptides

[0318] SEQ Sequence

[0319] ID NO:

[0320] 144 LIREEVQRRLEEIKQKIKEGKLDPFEAMSLLAELYELVEKLGDPELRKELEKVMEIVMRLG

[0321]

[0322] 145 SKEELKERAIRLLKEGDPFEAAMLLMHLAFLTGDPRLEEVIRLLEEAFELGDPELLKEI IKLLEEDVMEEAFTVWFQLENLAFRVDDPEQRKTVDTLSKLLQEAADRGDRELLKKVIKTARDVI SKVNS SELLERILRLLKEGNPEEAEEVARELLERTGDPRVEALVSLIEELLRTGDPFYELFVEMILRELR DEELLELVYEAVEKNDPRLLFEAWMILASLLDKTGDPKIEELLRLLQLVDRGDPDARRRIKELFK DEELMELVYEAVEKNDPELLFEAWMELASLLDETGDPKIEEALGLLQQVDGGNPDAGRRIKELFK DPFTAMELVARVFELALEKNDEELQKEAETLMEIVIKAAQNNDEEKVKEVIKRAKELLSKS DPFTAMELVARVFELALEKNDEELQKEAETLMEIVIKAAQNNDEEAVKEVIKRAKELLSKS DPFTAMELVARVFELAVEKNDEELAKEAETLMEIVMKAAENNDEEAVKEVIKRAKELLSKS DPFTAMELVARVFELAVEKNDEELAKEAETLMEIVTKAAQNNDEEAVKEVIKRAKELLSKS DDHAQAQLVKKLLKLWQDQGDDFYALLAVEAAANERVKKYLKKYYPEVYTLLEQVKRTLEKKRS SEEVEEAVELFKEDPKTALNLLFELFMRLVKEGDPEKAKLVQEVAELLMNGDPDKAREVARRVRK DEEVDRAKELAEELGDPELAELIEELKELSKRGDPRAVEWLRAMELANRLGDPEIQEVLIKLAE DEESETIKKKLDDILRWQSVNDPELESWERAVKDAKKALDNGDKNTARFLLKMADLARQFAS DKSERDIVEKIFKLVMEIMQYNPELAKKVHSIVEKAEHNQDPKLLKKALDVAEKALKTVSS DEVFKAAMKVLHLAFKKGDEEAMKEVQKVSELLENGQASKEQAKKILESLARRLKS DDSQVKDVESKAREHKKKHPQVSFLAFFVIEVARFAKEFNDPLALKVAEELLKEVEKRAK TEEEFRVMMLLMTLSPELNELVEELSEAMDRNDPELEKRVLKRLLELARELGDEEWRILEKLLK SMEDTVREALKAAKKTGDEFYVSMAEMAAQLIGDEELKQRVKEAERYVREY DESEEVKKMLEDLLQKVEKATDPTDRQFAIFMIELVLEAIPDPNQQKLAKTVLELARKLD DLEEIVREYVERTKDPIAREALELLEEIKKEDPETAKVLAEAWDAIERNDPESLKFVLLLIKIV SELLEEALELVRRGDPRVFEVLFEIWMLAVQKGDPELEELIRKVDELLQEGNPEEASKLLEKLLK DPFTAMELVARVFELAMEKNDEELQKEAETLMEIVLEAAKNNDEEKVKEVIKRAKELLSKS DEDAKMSIKLVLEIARDLGDEDVYKKVKSLLETAEKQSRNASSEEEKSRIWQDAARKVMKLFF DPFTAMELVARVFELALEKNDEELQKEAETLMEIVIKAAENNDEEAVKEVIKRAKELLSKS SEVEELVELWELIKKTDPELARDVKETLEELLKRDPKEVLLYLEFLAHFLGDPELLEKVRRLLK DKEVLEEVLELVEEGNPFEAFFLLLELAFKLKDPEIDEVAQLIVEAFDHGDPDLIRELVRKLLEK DDLDEIFTLSMLAMELAYKVGDPSLETEVQQWDTASELVDKDPRFREEAKKILKKLVEKAKKT NEEEKKKAQELVKQIEKILKQIDDPFYKEMVRMLLHIAKEMQDPEIIKEAYEAAKELLELAKS DPESDAKRIATELATLYKKLDPEESRKVADTAFTIAMRLLSSPDPNDQKKAQIIAEVFDKLAT DVKKLAKTVKEILKEAQKRHPTPEVKHEAEFISWFLDAATSVPDEKYATSILTELEKAARSLS SELFELVMEAMEIANRLGDEDLMRLLVELVEKALESPEVRELLEKLARLIIEDPEKARELVKKLK SFEELLRELVRRAGISPDDIDFVLWFVKVALENGNPEIAKELVEELLRKAGVPEEEIEKVKRLVE DPSEEAQKLVDKVKEDVKKKNDEDAEEQVKVAEELLKEAKNAQDPRDREFLVLLIKLIVKNLS HLEELNRHLLHLAFEAYFKIDDEHEAERFHKISEELQRLFENARSEEEKEKIVRLQEELIRRVI DEFEKHMAQLAVEAIEEILKDIDDPHYKDLVRKLLETAKKEQDPTKISKALEKAIKLAEKAKS RQEKLRRLLEKVRKLVEKGKLDPFQIMSLAAEVIQYAEKVGDETVRELAERLIEEWKLAQ DETKKVIDKLVSEAKKLADSIDDEFKRKLAQMAVESLEFEVNNAKNPEQVKKLWKEIHKIITDL SEKEREVKELLEEISRRDPRISFYLFFVLIALEAGNPELAEELIEDLIRRFGDPLLRELLELIRK DELEFLVWFAQVALEMVQEDPTNEEAKKVATDIVRDLDELEKKSRDPRAQELARRIKELYKKAC SAFKLALKAMHIANELGDEKAMKTVQDIVRLFDEGRISEEEAEKALKRLIKKLK SEESEKVKRWVETELTKANKQNDVFKAMSIVATALELAEELNDEELEKWVRSLAEEVFTKAV SLLERIEEALRRNDPFLVAMLLMNLAFMTGDPRVDEVLRRFVKAFEEGDPEKAKELVKELLKRVK MEASESAKELIKIVKSLYSNGDHNAKFYASMLKMLAEALELNAETEEIKKWKEAKSVADSVLS DEDKKTLDSLVSYVETVIQQANDPFLSFMLEVLLRFAQENNDPEAAKEAYKLAQDVKKNVD DTEEIRNELFKIAMIALRVNNEELYEVAQKAAELLKEVSKDPRKREELKKLLKRARELSKRS TELEFLEEMVRLVLENALENGLLSPDDVKKYEEKVKELRKRGDKDEVQKILEELLERLERW

[0323]

[0324] GEFYIELVEMALDFVRMLSGDEEFIKRVRKRLKEVKERGDDSKLQEILEEVSEYIERVADPFTAMELVARVFELALKKNDEELQKEAETLMEIVMKAAQNNDEEKVKEVIKRAKELLSKS SDDEFWRWMVKAAVDAAKELNPDPTVQTKADEILKKLEEALKKKDPTLVTRVLEEATRLAEEA DEKEELEKRLREAEKLVRQIQNEFYKMLADMLTKTFKEALDNNDTDQLRMIVEVIEELARDALS SDFLIHFAKEALEEARESGLLTPDEAEKWKKLLEEVRKRGDDNQIRKIYEEIIELVERLNG DDEEVKTWEKLAKEAINRIPDKDVRTEVEKMVRDLIKAAHSPDPQEVRFALEMVRMIIETFRG NEESEKLSRHLLELVERLVDDEDWKTARELAREAIELSNSSDHRDRERAEFLNLLLKLLT SEEDKIRKKVEEYVKKAIKRGINPSEVADWVAKYLTKAVREGKLSPEAARKIVNLAFELALKLI SEVEEIIEELRRLGNPEVAEILELALRVAKELNDPQTEDSITELARLLKEGDPFAKHMLELIARD DDSEEITKLLDDLVTKAKKKIKDPRDQKIAEDLAKEAKKLASSPDPFDKHMAKLLVEALQQFV DPSEEAQKLVDKVKEDVKKKNDEDAEEWKIAEELLKDAKNAQDPRNRSFLVLMIKMIVKNLS PEEREKLEELIKKDPSEAVFLALSILARLFEETGDPRIEELFKLLFEALERGDKDLLERLLKRIG KEEAKLNADFLVWMATVAARDSSPESRTIAKELIKEAKKLAKEANDPNVEKKVRSVEELIKKLG SAFELALKAMEIANRLGDEKAEKTVKDIARLVSEGRISEEEAEKALKRLIKKLK NEEEKHNAEFLLWFAKVMADYISDDDKRKKAQSLVEKARKLDIPEEAKKVYKIVSTILERDAK DLIEKVLEALERGDPELLHRLLFELVMKLLSLGDPRLLELAQKLANAFDDNDEELLRKLLELLEP SPKEELDKWSKIVKEIARKKNDEELYRIAERIYKDAKEALEKNDETDVKFLIEMLKMLSENG PAAETLEEAVEEGNERHVEFYLFFVLQILEHDPDSPEAKELRRVLEELRRKDPRVEELLKEAKRR DAEEILRELAKQVGVPPEEVEEALEILTRVLGDPKEAAFELLAILAEAAEKKDPRLFRELVERVR DEEEVRRVLFYAWFYLVHLDTEEARKLAKTLDDLWKLAQSIPDPKQKKEVLDKLKEFAERWS DPSEEAQKLVDKVKEDVKKKNDEDAEEFVKVAEELLKQAKNAQDPHERRFLVEMIKMIVDNLS DEEEIKRILEEEKRALEEAGLDEEAVRELLEESKREWDEVLRRNREEEWEKLKEVQRFRLAIIK DPELEEAFKLTMLLMHLSMQVPDPEQQKTVETLAKLLTEAADKNDKDTLRKVKKKAKDVYSKLS SEAEKWKELLEDAKKLVENKQASPEDIEFIEFFVNIAAEANGDEELLKKVKKVIKELKDKS SVEELIKELRELGDPELLRLAEEVLKLARKLNDPFLEKSIEELIRRAKKGDPFAKEMLEMLVHEG DKEIEDKAKEVSKRAKELVKKDPFEAMSLVAKLFEEAFRKNDEELLRWAEKVWEEVRKYIE DESLSKLVFTLLSLLAEAWKYDPDSAETAVELLIKAANNNDEEQLKKAVDYAKKWDRATS SLEEEARKLAEKLKRQGDEMAKELAKYLEDAVKNGDKNDVSFAIWMVRVFQE SVREAVEIILKRVGVPPEFIRELKEELEEAEKEDPREAEELAEFIIAFALNVPPEEARRLLEELQ DEEAVKELEKLAKELGDPELLEALKKLRERLEQGDPEANDLLFRLWFAWNNDPDLVKELVKRVP SVEELLELLERLLKETGDPDVSFILWFVRVALESGNPRIAIEIVLQAAEQLGDEELKKLAEELLR DEHEKEFLKALAKMALDLLRNASDPNLAKELVSTVKDLADLVNDEDAKELKKQADEVSSKAS TEKEKWKDKIFQLIMELLQLGLDPEEILKVAEKALELEKKGVSPSEIKKWVEKWKEVKKRI TDELKKEIEEVEKKVNTSLSKNPFLLWFARMAIEQAKEAVKKGDEEEAKSALTTVKELQKLVNS SEEIERLVRRAAKGDEEAERLIEEIIEELKKRDEFRAAMVKMILEALKEVPDSEELLEDLTRMLE DLEELLRRLREELGDPRDAIFVLLILLMEEGRLTPELLELADEADELIKRGDPRLEEVLKLLVKP SEMKEWRALVEEAEQRIREGKISPHEVRFYLEMARMAVEALGDPEAKKLVERLERLWRKIM SEELREIVKRALEGDPEAEKLLEEILKELEERGDPRAFLLWFLIVAIRFGNPEWEDVLEDLLKR DPELEEAFKLTMLLMHLTMQVPDPEQQKTVETLARLLSEAADHNDKDTLRKVKKKAKDVYSKLS PKEVAEVARLLVEDPNNKEALSLLMKLWFEAVERGDPELERLVRLVNELLQKDPREVRKLVERLR GEFYIMLVEMALQAVRENSGDEEFIKRVRKRLKEVKERGDDSKLEEILREVTEYVQRW SEEEVEELLERLKKISPDVQFYLWFLEVALRSGNPEVAIEIAEDLLRLLGDPLVREIIKLLRKIR

[0325]

[0326] DDEEVKTWEKLAKEAINRIPDKDVRTEVEKMVRTLIKAAHSPDPQEVEFALEMVRMIIETFRGSEEEEIEELVRKDPIQAYFLLLELANKLGNEELVEEVAERVQELYENGNPEEATKLLVELLERLR DPESDAKRIATELATLAKKLDPEESKKVFDTAFTIAMRLLRSEDPNDQKKAQIIVEVFDKLAI DDI QKEAKKVI EQLQRI KKT YPDNEKI LFLVELAI EAAQFAI KFNDEEALKVAKELLEAVKKI C SKEEVFKLVFKVMEVAVRVGDPTAESIAKTLVEQANEAALNNDDEKLKKVSKRLKSFIKTF PDELVEEWKKFGISPFIRFFLRIWEQLKRGEIDPEDAEKTLRDLLETVGISPEEIEEILRRLK DESLAKLVFTLMSLLAEAWKYDPDSANTAFELLMKAAKNNDEEQLKKAVDYAKKWDRVLS TVMDEAFKVTFEVMELADKVPDEKVSRLLKTLAKLLTEAADRNDEELLRKLIKKVKEVRKQAKS SEMKEWRELVEDAEQRIREGKISPEEVEFLLLMARMWEELGDPEAKKLVERLERLWRKIM RKEEAEDTAFKLFMLWLTLTNPEERKKVEKQIETLVKKLSNAEDPQEQKVAKTVKETLDLLRS SAFELALKAMEIANKKGDEKAEKTVQDIVRLVMEGKAPEEYAEKALKRLIKKLK DELEAVREYAKFLAEIAGVPSVSVRVSNGRIQVTLKGTDEAARAVQKILTEAVKHWGVDVDVQVR SDDDVKKLLDELEKLADQIPDEFVSLMVKMIVDAARKALKLGDTEEAKMIAKVAEELVSKG SVEELLELLERLLKETGDPNVEFILWFVRVALENGNPRVAIEWLEAAEQLGDEELKKLAEELLR D VD EAVKAVE EAVRMVQ KH P D D EAL RW FARWI E FVKE LAS S I E D E DQ S KT AQ KRAT E L E RKL E DTFTYLIKLLSKGDPFARHMLENLLEDAKLRGDEELYRLIKEAIELVEEGNPEEAEKLAKRVDEK DAREIFRVLALAAETIEKASEEEVRRELEKLIEELFQAAHRNDEEALKKVLKRLKELIKEQK DEFELAFTVMSLAFQADLAEDEKELRTLQSLIEEILRKAQSEFLRKLAEDLLKEVRNNS DEEETIRSVLKELKDDASNASSDEDREFLIWMVNVFAETVPDPTARKIAKEINETLSKLS DALERAKELAERLGDPLVEVLLYFLEQALRQTGDPKYEKLAELILELLERGVPPEEVLHLVEELG DIFELMFLIMHLLKDNPELSELFREVLERLWRGEISPEEALRLLKELAREAGNPEVEELLKRVEK DEEEVIEKWRLYEEGNTFEVMHLLAKLFEETGDPRYQELLDLVFEVIKKGLSPEELEKLLRKVP SEEEELLKELKKRVSPEAREYVERLLELLERGDPNFLFVLLELLFLADELGDEELKRIVEKLLKK SEAKEWRALVEEAEQRIREGKISPEEVEFLLDMARMAVENLGDPEAKKLVERLERLWRKIM DEEEVFKLFFKLEEFAYRQGSEEAKKALKKFHELIEKAQHVPDPTERKKLLDSASKVAKSVIS SLELLEELVELAKKGDPRVFELALEAMQLANERNDPELSELVQRIVHLITNGNPEEAEKLVEKVK DEVKEVAEKAVKTGDPKLIKEALELLTKLVLKTGDPELRRLNTLLFEAVMVGDPELLKKILKEIR DWRELTETLRRLVEKAKKSVEDPFVRALIEIALEAAEEASKRKDEELLHLLAEWEELRHAS SVFELSMKVMHLVAEVERNAEDPEVQKILKEVNELVREAAEKNNEELLKRAEKLVKKAKELVS DDEEAKKLLDLARKALKEVNKTASPEEQKQLTDAVREAEKLLHSPDPFQREMARMLLEMWMLG DDENVKKDVKTVKDIAKKLNDERILKTLQDAERKAKENNDSRIVRDVAFKLWFQLMNVE DADSFYSWLLEEALEQAKELQDPEVARMAYESAKDALEKYPDSELVRKVLKKAEELQRKLTS DESLFEALLELMQVSPEAQILVEIISDLFVRGVPLEEIEKIIEEEAKKLGIPEEVLRLAKELLRK SELSEEALRLLEEGNPREALKVLFELWFELVDRGDKELSELVQEIAELLIRGDPDEARKLIKELT DEVEKLTTEAERVATKLERSPDPDNQFVAFMIKMAIEFALTISDKDAAKEALKSAVEMARTY DEEELKELAEKLKEDPQVAVYIWALLAHLWYKTGDPEVEKVERRFEELVHRGDLETASELVKRLG SLDEVEEELKKLVNDPEAERNIEFWWMAKVANLSPEELLKLLEDSARASGDEELLRAVKELLRK DDKTLTEVWQHASTLAKQKNDEFVLHFIEVMYTTAQELGDPDFKRNVEKVLRDLSKDLEG SEELREELKRAVDRNDVFLWMLLLDLSLETGDPRYEELARRIVEVDMRGDPETLRELLKLAKEP EEDELRKLVEELEEKARKVITSDEEREIVERIIDDAKKALKNGDKTTVKFLAKMVEMFIRTG

[0327]

[0328] DEEVKEILREALNLDPEEINDLIFRLWFLAVQLGDPELERVLDEIAELLQKDPEKARKLIKELLKPEELVKEVEFLILMLKVIATDPEIRKDIERLEKEIRTRIKKDPRFAEEAVRRLRKILKRLVEEI DEYQEKANKLYKEAEKLVKTYRSNELVKELFEIATRLLKEAREKQDPMLFAFVSMAIDALKENG DFTKIAKELVDSARDLRKQNTEFAKHMVKLLSEEAKQLKSNIDDPTERKWSEAIEEIDKLSKG DKEIAELLKLLLEDPNDKEVADRLFRLLMELMHRGDPLLELVLEAAQLIEEGDPRVREVIKKVEK PEEEEDVKRIIEDLLKLIQNASSEHEANFLYHMVELWEAVNSEYAKKAHELAKKVLKELRKR DEEHYRMNIDFLIWMAQTAVSLDPDVLKKITKKAEKIKKEVPDPDQWKLLEKLVQEAVELAN DDEEVKTWEKLAKEAINRIPDKDVRTEVEKSVRELIKRAHSPDPREVEFALSFVRMIIENFRG DESKKIADEVKKTAEKLIKNGNVFEAMSLLATVLEKALKNNDEKLYKLVMTVAKEVFSLAKS DEEFATWFVRIALENGDVELAKELAEDALKQGEISPDAADRLRKEIESVEKR DSEEVKDLSKRLQEAAEKAQDPNMLDKAFKVWFQLVEAYNNNDEDKVKDASKKAKEVIKTIDS DEREELAQEVAEEMIREAKHAESERHAEFYLFFAERLARAINSEETLELIKEVAREIREKG SSEDWRELLKAAKKTGDEFYVLLAEMAARWIGDEELKQRVKEAERYVREY SEEEVREFVERTGDPDVKLALELIEELKRRGDPESAKIVTEMVLKLIRTGDPFLRALLRMMIENG DEMKKVSEELKKDIKEALDKNDSHNLRFLIMIAKMVASTTNDEDLKKFAEKAEKILRDRES DHEEEIRSVLKDLKDRASNASSDEEREFLIWFVNVFAETVPDPTARKIAKEINETLSKLS RSEEDLRFRLHMIELLLRAIGVSEDEIKKINKTLEELIRKGDSETFKRIEDEVEKRVKKFT DDKTLTEVWQHASTLAKQKNDEFVLWMIEVAYEAAQIYGDPHVKKEVEKSLRDLSKKLEG SDEDVRSLLERIDSELKNVSNEFVKFFVELAVREAKEALKQGDKEYAEMVLKAAQEVLSVE DEEWELLKKLVNDPELEKVLEILLKLIKEGDPDAQFLLFMLKLALEVGDPDTVRSILEDILRQT DEAFEAAHILAESLFIAREIGDEELIRLAEHLLELLSKAAEENDKDLVREILRLARESLKRSR SEELREELKRAVDRNDVFLWMLLMTLSFETGDPRYDELAQRILHVASEGDPETLRELLKLAKEP SEELEEVLELLKRDPKKALELLFELWFEALRRGDPELLRVLEELNHLILEGDLEEAKELLKELLK SETEFVKFFVDVALESAKELQDPHVAEIAVRIAEDVLKEDPTDETVRELQKKAETVRDQLKS TVEELARKLLEEGRLSPEEIVEVLLRELRKLGIPPFEAALLLLKVANDLNDPELYKAVQELAERT PQLDKMVKDAQDLADKIVKKASNPEIQKKAKTVKKLLEEARQKKDEQTVAELAFKAMELWFMVT SADEATDSAAKAAEKGDSEYIERLLKKAKSDNIIDEFHIWFIKMLLEFARE SEETITRLYKDAKKKAEDKKNEELKKTVKIAKDLLDTAKNVDDRMTKEFLETMAELALRDV DPELEEAFKLTFLLLHLLMQVPDPEQQKTVETLAKLLTEAADKNDKDTLRKVKKKAKDVYSKLS DPQKELHKVYETAVKRIRSIPDKEKQKEWKEATELLRRIAKDEQEKLSAEFLSWMLKVLS DPEQELRTLEKIVTEAAKDPRRSEDFFKAASRLAELIYKANDPKIQTIAREVLTKAFSIAN TVEELEKLVEKLTEKVKRKDPRLSEKAQRILDQAKDLLSKAENPLLQEMIARFAVSQLQRLV SMEDIVRELLKAAKKTGDEFYVMMAEMVAREIGDEELKQRVKEAERYVREY DKSDAQRILFEAWFLWELGDEELEKEVDRLQKEFLEADKKNDKDLLDEITRQAEELLKRIKS DLEKLKKDAKKALEILKKLNNEFLVWFLENLVKALEMAIRKNDEHWKEVSKAIQEIIERLQK DEEELKKVAETAKTVAKQKNNEEVLKAIKEMATEAEKLIKSSNPRDQMNAEFIIWMLRVLIED NEQEKRNASFLLEMAKMLADFISNDDLRKKAQSLVEKARKIDDPNVAKKVYKIVSTIFERWK FEDAEFILWFAKALVESVGDEDALEKLKEFEKKVTEAIKNNDEEKVRKLLSKAEELAERVLQ DEHLKKLEKKVRTLLEKARKNNDEEVMFLLFFAELSIEMVKMNPEAKEDAQKTVEDLLKQAEST DEESSRAAKDLLKKAKEAVERNDSQTVSFLLWMVKAVAQNAGDEEVEKSVSELEKLWKKIS DEVEKLLREANELAFKLLSDDWRAFDLLQTLARIADEANLSGDKETASKLLKTIIELLKRG DPSEEAQKLVDKVKEDVKKKNDEDAEEAVKIAEELLKAAKNAQDPRDREFLVLIIKMMVENLS SEEEYKKKIKELLRRGDAFEAALLAMEAFFRLGDPRFEEILRIIHELFERGISPEEIVEILERLE DWRELTETLRRLVEKAKKSVEDPFVRWMIEALLDAAEFASKNKDEEMLKLIAEAVEDLRHLS TEELIKLVKELAKRNPEEAKKWEELVKLLGLDPLAERMLKELVEEYYRTGDPFYLFMFEVLVRN DELEKWKELSTDADKAVKNGDKDKVKFLVWFLKSWEFDPDNPAAKLAKKVIEKLEKLIKS

[0329]

[0330] SEEIKRLVELVKKGDPEALKRLRELLEELKKKGDPRAFLLWFVIVALEAGNPDIAKEVAEQIERRDPSEEAQKLVDKVKEDVKKKNDEDAEEEVKIAEELLKQAKNAQDPTEREFLVEMIKMLVKQLS QEEYKRLLEELLRKAQKAVKNNDQHEVSFLAWFITISAENVEDEDVSKHFLELAKELHKFVNS DRDEQLKKAIKDLLEQLKKASSRHEASFLIWFIRLLREDAQTPEVRTLATEADKIYKKLT DEWQELSKIVKSLIDELKKSPDPKDQFAAAMAEMAWEMAERAQDPNVRREALKTAIEMVRDT DPFTAMSLVARVFKLALEKNDEELQKEAETLMEIVIKAAQNNDEEAVKEVIKRAKELLSKS DDTKKLHDTVTKYAQEVLKRARDEDVKKVIEELLRDAQEVIKNHDEETAKFLYLMIQLASKQAI TEDYEKVKRELEKVEEEAKRKNNLEALFLVFMAKLSLEAFNLDPDNKDQIISVAKDLLKLAKNS SAEEALDSAVKAAEKGDSEYIERLLKKAKSDNIIDEFHIHMIKLLLEEARR SVFELALKAMEAANRKGDEDAMRKIIKIVQLASDGKIDESQASTRLRRILKKLRS SALEEAERLARELGDEDLLFLLAMVKMARSTNPEVAKELLEEAVKLAEELGDPDLLKLLRLALKE TFEERVKEALERNDPDLVKNIIFEAVMELLKRDTEKAQRVAELAVELLKEGDPDKLRELARRADK DDETKELQNEAFKAVMIVMQLNDPRLEKWRDVQTRFEEAQKHNDEETSKKVLKKLVEIVRKT SEEKEWRHLVEDAEQRIREGKISPEEVEFLLMLARMWENLGDPEAKKLVERLERLWRKIM DEEEKKEKTLDKLLSEAKKKSSQNPEVQFLAWMLEMLVEQFKLVPDPEIRKEIYKAASDLYKQT AELAFKAMSLAAQLAKALLDGDEDKVKTARSLLDKVEKEVKKQNDEEVLKFVQSLKKTAEDVT NEDEKRNAEFLLWFAKVLADSISDDDGRKKAQSLVEKARKLDIPEVAKKVYKIVKTIFERDVK DVFTVALKVIEILNRLINNAEDERIKKLLETVSRLWEAAERNDEKLVRDLYKTLEKLVRELRS DESEKFQTWKELEDLVHSVEDPTRRKKARNIAFKLVMKLMNAEDPKEFKETVDKAKEIVKKAA PLEEQLRELLEKAFEAWAKIRDPEEAETFVEKFQTLEKQFRNASSEEEKKKWEKIKKLIDSVL SEIEEVLEKLKRLSPEISFFVWFIEVALRNGNPEVALKLARDVLERFGDPLVKELLELIEKLVRK DDKERLRKLVDDLIRKAKEVKDPETVRNLLFQAIMAAMLINDPELEKKVKEVFKEIKKKVS DEEEKLQTVIKELEDLVKSVEDPTRRKKAHEIAFKLVMKLLQAEDPKEFKETVDKAKEIVKKAA DIDEKLDSLVKKAKRYVDETNSDELKEEVSIAEELVRRAKNASDPNDKRFLTWMVEAIYELVI DPKKLDELRKKIETLLRKVTDPFAKEAVKAALELAEEALKKQDPFLEFFARMAYEFAKEMQ SEESEKVKRWVETELTKANKQNDYRKAINIVFTAAMLAMQLNDEELEKWVKSLAKEVMSKKR D P KE E L E RVT KT AQ KTVTELNDE FAL FMVNN L VE FAE EAVRNN DEE LAKT VI RALQ ET F KNMM GEFYIMMVELALQTVRELSGDEEFIKRVRKRLKEVKERGDDSKLEEISREWEYIERVL DLEKEAKTWTLLKRLKKQVTNPDVKKLVDEVAKSVETAVKKKDEDKIKSLVFEAQILI FAI DLHKEAFKVWFEAVELIFKVQDPHIRKLLEHLAKILQEAANKNDEELIKKILKLVQKVIKEARS SIEDIAHKIISLALELGDFDALSQAAEVLEKWLSGKISPEQAKKLADRIRKILE DPFTAMELVARVMELAIEKNDEELQKEARTLMEIVLKAARNNDEEAVKEVIKRAKELLSKS TEEEKKAKKWVEKLVEKAGLDDFYKEMAKLVFEDAVLQGVSTKEARKIVKKVIKEARNV DKREEVHTLLEILAEAAEKRDEETIRKVASRAQKLAKEIGDEFLLSFIQMLFEYAQQLLS GELAKRALELIERGDLDEVFKLWFEIVERANRTKDPELEELLRLIHLAAQGDPEAKKELEKIIRK SELLEEAFEIWFKWEIAYKLNDPDKQRLAEEVTRLIETAMNVPDPTQVKKLVTKAKEI FKKVK DEEELLREWERFKDDPFHVAFLLLEIAERRNDPELAELISEVIEEVSQKGLPPEELLRLVLELK DFEELLEEARREGDPRTLVFLIALLALQRGDPELVREIEEVEELIHRGELPPEELRRLAEELIKK SVELLFEVWFLIAQLGNEEVLRLLEIFAEAAIRGDPDLLEKVERRIEELSKTDPAARLVLEAIRG DEFLIMLAKMLLEFARNNGLLTPDEAEKWKKLLEEVRKRGDDNQIRKIYEEIVELVERLAG SEETITRLYKDAKKKAEDKKDEELKKAVKDAKDLLDTAKNVDDEMEKKFLEWFAEVLLRAV DIDEKLDSLVKKAKRYVDETNSDELKEEVSVAEDLVRRAKNASDPNDKRFLTFMVELIYELVI DLEEKAKEAYEEGDLELLVFLVLMILMEKGDPRLEELLKLAEEVKKTNDPELRKELRRLAYELIP SADEASDSAAKAAEKGDSEYIERLLKKAKSDNIIDEFHIWFIKMLLEFARE SDSDEISSAVETLLKQAKDAPDPTDAEMLLWFAKVLVDYLNNDELRKKVEKASKEVEKINRS DESKKIADEVKKTAEKLIKNGLVFEAMSLLATVLEKALKNNDEKLYKLVMTVAKEVFSLAKS NEKEKMDAQFLLWFAKVMADNISDDDKRKKAQSLVEKARKLDIPEEAKKVYKIVATILERDQK

[0331]

[0332] DPESDAKRIATELATLAKKLDPEESRKVFDTAFTIAMRLLSSSDPNDQKKAQIIAEVFNKLAT368 SSELRDRLKKTFQELLKRGVSPWQAFEIIEKLLEELFRKGKLDVDVAFEVMHILAELMRKWE 369 DRTEKDQTVIKELMDLVKSVEDPTKRKKANDIAFKLVMKLLKAEDPKEWKEIVDKAKEIVKKAG 370 NEEVKKEIKSLEKTVSTLASKDPFALFMAQVLIEFAKEGLKKGDEEDVKEANSIVRDIIKRLQS 371 DPKKLDELRKKIETLLRKVTDPDAKEIVKDALELAEEALKKQDPFYEWAARVLYELAKQLQ 372 DDEEVKTWEKLAKEAINRIPDKDVRTEVEKMVRTLIKRAHSPDPNEVEFALLMVALIIETFRG 373 NEEEKKKAKKLLDEAKKLADQISNDDLRKDAQSNVERARKIDDPRDAKFVYFFVKLIYEFVQS 374 DETQKRQVEKYVRTIEKLLKDAKDPKEITELLEKALTYLERVNSELAMEAMYRLMEVYFEAT 375 NEELLTKFVLEFAQSLAQRASSQEEVDKISKEAQKILKELNVDPRLVEKVSRLVQEYYKKS 376 NEEEKRHAEFLLEFAKMLADNISDDDGRKKAQSLVEKARKLDIPEVAKKVYKIVTTIWHRDQK 377 PKEELEKLVRTLEELVKKKVRDPFVKWMADVTVTFAREAVKSGDKDLATMVIEDLKELLKTI 378 DIMSLAFEAMELANKNGDEDAMKTIQKLFELFDEGKIDPTQVQRILRKIAKRLKG

[0333] 379

[0334] SEVEELLEKAKRTGDPRLASRAIFLVLMELLKRDPEEVQRVQEELLRLLKEGDLEELERLLRKVQ

[0335] 380

[0336] SEVLKELKRLLEEGDPEEALRLAERLLKEAKEKGDEELAFILWFVIVALRQGNPEAIRVARELVR

[0337] 381

[0338] DKEELKEELEKLIGDPDAAELLLHYIEAVRRGEDPPERVEALVEFIVKIAGLPPEVEKRLRELAR

[0339] 382 PHVENLVFTLVMKAMEAINKNDEDTLKKLKDTLDELVKKDPRFTEAAKLAKKLIKEQEELT 383 DPVEFLSWMAETAISLLEELGLLSKEELQKWKEIVRKLTERGDTENLEKVLDKITEIIEKLQKS 384 KEEVNNQAFKLVMEAMELANSSDPNNRTKAKELIKEAKKLAKEVNNPEVEDTVREVEESIKKVG 385 DEFTEIVKELVKLAEEAVKKNDEDSVKFIEAMLKMMKEAATDPKQRELADRAIKKVQKLLKS 386 DEHLKKLEKKVRTLLEKARKNNDMKVAFLLFMAELDIELVKLNPEAKEEAQKSVEELLKQAEST 387 DPFTAMELVARVLQLAIEKNDEELQKEAHTLMEIVIKAAKNNDEEAVKEVIKRAKELLSKS 388

[0340] DEVERWELLKRLGDPVARELAELIEEILKRNEEMARSAASSAEMALKKNDPESLKFLITLLKMV

[0341] 389 DIDEKLDSLVKKAKRYVDETNSDELKHAVSSAEELVRRAKNASDPNDKRFLTAMVEMLYDFVI 390 DDSEEITKLLDDLVTKAKKKIKDPQLQLKAEVLAWFAKVAASSPDPEAKKVAKELVKELQKLV 391 DPKKLDELRKKIETLLRKVTDPDAKEAVKMALELAEEALKKQDPFYEWFARVAYEWAKELQ 392 SDEDEIFTLAMLAMEAIFKAAKTEEEFERLMKKLQDVIEKAQQLDPSQQKKLLKEVLDEVKKL 393 NEVFELALKALHVAFKVGDEDALKLIQKIAEKVSDGKLDEKQASKLLKSLIKKLE

[0342] 394 DVDTEALRAAEDMIELLKNAKDPTEAKFAKTMVELLVELVKDPEARKIIQKKLKEAEEYVNKL 395 GEFYIEMVEMALDMVRMLSGDEEFIKRVRKRLKEVKERGDDSKLEEILREVTEYVERVL 396 DAFDALSILAEAAELVRKLGDEELKKEVDRLLEEFTEAARNNDKDLLDEITRQAEELLKRIKS 397 DDDSYKKKVEDTLESLIKKAKDPLARKVIEELARDAKKAVDNGDKEKVEFLVLLAKLVADNAAS 398 TDAKEIIKDLKKRFEEAVRRGVDPRQAEFLVKMVEMLAEDVGDEEVKEWKELRKRLKELKT 399 DELEKMVKRISTEADKAVKNGDKHEVEFLVFFLKLWEDDPSNPAAKLAKKVIEKLEKLIKS 400

[0343] SVSELLSEAASKNDPRLLEEAVELLVKELGLSPEEEERLRRLVREALKRGTLFEWMELMTLANR

[0344] 401 DPREAEELAKKVEEVASKRGDEVMLRAVEEIKKLIETAKKSPDEFYRSFVDMASDFLKEITS 402 DELEKWKRLSTEADKAVKNGDKHEVEFLVWMLKAWDFDPDNPAAKLAKKVIEKLEKLIKS 403 SEEKEWRALVEEAEQRIREGKISPHEVEFYLWFAAVAVELLGDPEAKKLVERLERLWRKIM 404

[0345] SLEEELKRLISERLGVPPEFVEEIVEFDSEELKVPPEEALLELAKAVARLFRDPELEELIRRLEK

[0346] 405 NNEAQFLLWMVQMLVEAAKLDPDSAEELSEAIKDLLKRIRKLGDEDASKKATELLKEAEKVIS 406

[0347] DEEEEKLEELIRRGNTEEAKKIVEKIVKELGLDPFEEWFLKLLVENAKVDPLVLEVLKRALKELR

[0348] 407 SKDEKLHDKLFKLWFLLVNLHTEEAKKLSKKVHTAAEKASTLSPEERRRLLEKVAKEAEKLIG

[0349]

[0350] 408 DESLAKLVFTLMSLLAEAFKYDPDSAETAMELLMKAARNNDEEQLKKAVDYAKKWDRVLS

[0351] Table 6. BMPR2 bindersSEQ BMPR2mb

[0352] ID NO:

[0353] 31 TEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWRILVQLVEAAKEAEEIN BMPR2_1_1_M2

[0354] 32 NLEALI FFNRVQARFLGSERYLEVANQAEEALRRGDREKAYQILLESEKSL

[0355]

[0356] Table 7. FGFR2 binding polypeptides

[0357] SEQ Sequence of target binding polypeptide

[0358] ID NO:

[0359] 11 DRRKEMDKVYRTAYKRITSTPDKEKRKEWKEATEQLRRIAKDEEEKKKAAYMISFLKTLG 410 SEELIKKALELLRQGNPDRAAQVLLFLAFQTGDPRVRELFSLLVEAMQKNDPELLKRVRRLLEET 411 LEKLERVALLAVRLYMRIGDPEIAKIWFKVMELLHAYQAGHLDEEEAKRRADKLEKELRKFI 412 DPSKELDKVYRTAFKRITSIPDKEKQKEWKEATELLRRIAKDEEEKKLASLISLFLKTLS 413 DRRKEMDKVYRTAFKRITSTPDKEKRKEWKEATEQLRRIAKDEEEKKKAAYMILFLKTLG 414 DKKELSDTAEKLLKDALDKNDDSKFISAVFFALKVAKDVNDERLERIWKLYNEFMNRYH 415 DYERVISRAIELALKKGDEKALHILNEIFFAAHRGELDPTEAERLARRIEKKLRS

[0360] 416 DLREKLQELWWRALEKGNEKAERLALRAFMGAWHGDTSEEEAKKIITEVERLIKS

[0361] 417 SEIHKKLARLLSKAIQKGDRKANHIVIRAAMAYEKGLIDPDQARKLAEKAERILRS

[0362] 418 DIEHLLARAIHRSAQLGDREAVELLTRIFFALEAGKVSEEWAERLARTILEHQRN

[0363] 419 SLVERVIEQVEKRGLPPEALVLTVWLALVKAGDPEKARLVEELAREVFEGKLPPEELKRLLREIE 420 DEVHHFLLRLFFKDPDDEDVERILALFLLLKKEGIPPEEVTEIVIKFAEKLGNPELAKELKKILK 421 SEEERIAKLIIELLKRGINPERAVARVMFEIMELGDPELNHLVLRFFFTLKRDPEKFEELLRKLG 422 SKEERLKELLRLIKEAIKRNLTEEVERLAREAAEIAQELRDEEVLQEALLLWFRFEIRRR 423 DELFQRVMELAFRLVLAAREKNDKDAEERAQRLFRSVAKALESGDQDTAKTLVDRIEKYVKKFG 424 DPEKIIRELVRLALEAARKLNDDRVATAVLTLLYAVLRLEKRDPEKAKKLLKEVEEKVKKLIS 425 DEVEELSKKVLHIATRLAFSPNPEDQQLWFKAMRLLDKAESIPDRDKATKVLRKAEKLVSSS 426 TPEEVMRTAFELLSEAARNNDRDRLKEIVKWVEDLVRKDPRVKDALEQVRLTLRFMTHLAS 427 SERTASEVIMHAMQLGDHHVMREANRVYFGIVRGELRPEDAKKLLDTLRRQLE

[0364] 428 SEDESLKESLKHASKFFDIPSFEVRNSNGRLSVTVDGDDEKARMFLITAQFAAMLRGLKLDVRTK 429 DLREILRKLIEISEELLRKATDEELRELLELVIALSKEALRENDEELLEAAIRYNEVLIRNA 430 DFDTHEFEELEKVLEEARRKNDFRLAERAARLAAESLERYRDAELVREALQEALLLWLALYK 431 PELDIAVWAQIMADKIVKKASNPEIQKKAKTVKKLLEEARQKKDKDTVQKLWWKAVTLWQLW 432 SERKLAHLLMIAVQQGDHKANVMLLRAVFLLHKGEASEEQADKIADEAKKRLS

[0365] 433 DWKEIEKTARDLLKKARENNDRRTAFLVAVALKLAEIARKIQDPEERKRIEEVAKAALADA 434 TEEKIIRLWLDAARKGDRKANTLALRILFGLKRGEIDESQADKLADKIKKSLS

[0366] 435 SRLEEIIRLAEELGNPYVAETLKMFLRAVRNGQLSPEWALANARALADYFGDPELLRLIEELIKK 436 NPYEELLRLIERLFERLQKENDEEVEKALSFVNALLMLIARTTDEETKKELLRLALEKAKKVL 437 DEEEEAREHIQRQLEELEKHDPRAAVIFAAYIIERARENNDETARKIAEKLLKEIEKRYRS 438 SEELKEEAKRLIKLLKDLARKASDRRERFILAALAASLEKALKNNDEELLRKLIEVAKSAIRRIQ 439 SREEVLHILNHLYFIAYERNDEELQHILARAMYIVFELLNRGDEESAKKVAEKAKKIVDKK 440 DKEELIEEAIEKLAPSPEAERYLRELLKEFKELGNPEAFLFLLWLAAERLGNPEVKRLVEELLRR 441 DDKEKTVKTIVRKLLRRAAHVDDPEVRTALLTLAYILERALENQDDEALKRASKLAEKVLKSG 442 HETRDLNIRIETKREEELEHAERAQRLYFEAYLDALRNRGNKVDAEIRLHIDGDTSHSRAHFRFT 443 SEEHEKAREIAKKLAKRVRSGKLDSRQAKKIVARRYHHASRSGDKELAAILILVLFLLDEILRS 444 DEAQELARIVFELIVRLHRSPDPKDRKAAHFALRLLMKALKAQDPNERRSLLKTAIKYVRNT 445 DITWHARYTFHRHDEQAKREAESLAFLEVYKLERRAEELGLTSRFRIRSHEHNNRITIHLEIHIH

[0367]

[0368] 446 DPEEIKQRAEEIKKEFQKKGVSPEIQFAIEQVIKYALEVGLSPKDIRTTIELTVRFAEELEK447 SPEHAERLAKMLETLVKHAVKNPEERKTAETLIKQVKELIKKGDEDSAQELLAKVYKIVWKAQH 448 DKDEWQLALRILFEAYRRGNEEYYRKAQHLAALAFEAAEKNDEELLKKIEKKLRTLWKKLQS 449 DDDKEKKKKVKKKLDSIVEQAKKDPKHAMSLVLSIWMEAHRKNDEIVAKIAFEAWLKIMRIYSK

[0369]

[0370] 450 DEHLRKLLEKVMTLATKAWENNDEKWSLLAEAQFWIALVDQDPRKKEEAQKRVETLLKQAEST Table 7A. FGFR2 binding polypeptides

[0371] SEQ ID Sequence

[0372] NO:

[0373] 451 SEKLFKDINSLVI SAFMAGVISEHEASRILRRLFEAYDRGLTEEVKKIKKEVKKLIKKARKS 452 SLLSQLLMEALQKGDPELNHLLLAVIFLAHERGISPEEIDKLLEKLERELGDPREAIKLLLELLR 453 GPLNLLELTLWLASRRNPVARVLLRILRELRKRIDPEEVLKILEEVARRLGDPFLLRNIEFILRL 454 SEEEKLTELSERVQKFFGLEHVEVRLNNGTITVTVRGDDERADQAIFFVRFFAQLAGIPVDYKKQ 455 SEVELATKMIKHLAKQFGVPSVSVSYSNGRVRVTFKGNDDRVQQARLTALFVAHLVGVPLEVQTR 456 DPDEILDRVAQTLLTLAFQHPDDPELLKAFRRWEISQKAKKLSDPTQKERLARLAQRILEEVS 457 SIEEEILRWQFLAFKLGDPTLARLALRASHAYIEGDKKKARELIKKARKWY

[0374] 458 SFEEVAELLRRAGLSPTEIITILYRLLRAFRHDPERAKELVEELVKKLGLPPEARELLKELLTKH 459 TKEILQKVIEIVLKLVKKGVDPEQAVKIIRKILSELIRQGKISKEEANMATVTAMFVAHLAKG 460 DEKTLAQEIIEEAKRAWEKYKDDPDFLREFLRELLREAYELRLEEAIHFIQFLFFRLQLEG 461 DEIVRELEKLAKLPDAHMRLTVFLIRLFIHDPNDEKVEKLLAVLLELLLRGVSPEEIIKILKRVE 462 SSEKLKRAVEEARRKGDFELLWRIQLIALELGDEEVQNLAARAVMEMAKR

[0375] 463 DPKEEAKKVTRLLLKLAHNADDDEIRVELLTLAYALRRALERNDEEALKRVISKARKFIKDVKS 464 SRDELVEENIRFLARSAGVPSVSVSVSNGRISVTVDGDDEAAQELSSQALLYAHLAGLDLKVTIR 465 DEEKEKLRELAHESIRVALKRARSDEDHLRIARLYLALEKAVENNDKDLVKHLLEITTELAKRL 466 DKSDAEKILREALRLVWELGDRELFHLVSRLYLAFVRADKNNDKDLLDEITRQAEELLKRIKS 467 NWEEAAALALESASRAGDRELNEEILRIFFAVRRGEISEEEAHEMLREIAEKSR

[0376] 468 SELLEKELRELLKNGNLEQAVLLIWFALHEAGDPERAEEVWKLLREALERGDPDLVRKIIEEALK 469 SWEELYEKAKKLVEQAKKENDEDKLNKATLLVYMAHLAASELGDPTAQTSVERLAAHVFHVAI 470 SEEIEELIKELFRKARNPEVRTVLATLWLALERARSPEEREEIERLLRLIEEDPRLAREILKKLN 471 DSKELKRLLKEAEKKGDLELANRVAFLAHELGDEEVLRLASEVLSRIMG

[0377] 472 SEIHKKLSELMMQAMQKGDEKALHIVNHAYFAYERGEIDPDQARKLAEEAERLLRS

[0378] 473 GEEVLEKVERLAGGDPRLIAYALLRLFREARRRGDKDLLKKIEEAVELLERKDPELARLIKNSLP 474 DEWDERLKETFKKLQELLKQGKEEEAEKILKELIHEAEERNDEEAVQKALLLWFRLLTERRK 475 DHWKEVEKWADTLLDEATKQNDPQKALFILFMAFLLLRRELSEEEAQRVISKLIRKVHERLR 476 DEEEEIRELIEKAVPDPELRRLLKENLKRLEEDGVPPEEILLTLAFAAERYGDPEVARLLFELLR 477 SERELFQRNVEFLARTAGVPSVEVRYTNGKYHVHLHGDNEALKDAKRQVELFAMFANLDVKVTTH 478 DETTKTAETLIKQLAEKAKNVPDPINVLTLLFAAMLLAKEANDPRLEKLVDKLVREVLQYV 479 SEEELRRLKELLKKVNSPEAQMVLVTLLYLEHLGVSPEMVLRLLEEMLKRINPEALKLVKELLRR 480 DRTEILTLAFAILLMAKEKQIPEIEKLARKWHLFSAVQRGELDETEATKRARHLWSLASSY 481 DEIQRELNHVYFEAHERGDEELVQKVTRVLYALFAGKIDKKEARKLVEKLKKLLS

[0379] 482 DEIRETHHLILRLFFWAEEKNDREAEKIAARLLELQAHALARRDEELLKKISKEAAKIVKELLK 483 DPDHAARLVNVLAFLAVEKNDEEAQREVSELIMIVMRAKENNDEEAVKEVEKRAKELLSKL 484 SEIEELLVHLLRLARELGNREALHWLRAIIAYKRGRADEKQIRKLLKKAKSLLS

[0380] 485 EVDELQKIAKKVRELAKKQNDEAALRTVEIALRALERAKQKNDEREVEAAKLAIAAAREHVEG 486 DQETVKKLESKADELAKKITDPRKRFLASWKVNARRAKNAEDPNDQTELIELAEIILRHLG 487 DQEEAEKILQRLLRIAYEARDEEFFLEVSLLAFAHELALEEGDHEALEESIRIAEELLKEA 488 DEELKTLEKIVARLFHHAARNNDRKSLELATRAFFALVEAKNADEEQQKKLKTLVKQLIKKIK 489 DLEEEADHLLSRVLLVAFERNDEEVNHIVLQLVFLLQKAKEENDEDKLKTLLKKAIEIARKLLS 490 SESEILKHSVSQAAKFLGVPHFEIHDNNGKFTVTVRGPDHVSRMLKLTVRFIAHQLGLDVKVTTQ 491 SEEETFKRLVQEWARKAGVPRVFVTTYNNKVYVLIHGNDESAKRLTKIVKKLAKEIGIDVKVQTH 492 KWEQIILFAKFLGNPEVAEIIEKLLRELKERGVSPEEIERIIEELLEELGNETLLRNFRYLLEQ 493 DDDKEKKKKVKKKLDSIVEQAKKNPNHAMVLVFRILEEAQKKNDEIVLKIAEEANHKILRIFFK 494 SEEDKEHVAMWVIFLATQAAHDEDLERKARKKLDKILKDLKVPEAEEFKKMFDEIVKQVRSMLK 495 SEIAEKLLRLAVKAYEKGDRKALRIIARAQIAYAKGEIDPDQARKLAEKAERILRS

[0381] 496 GEELEEIVRELRKRGISPIQILFILMEILFEKGDPVAHLLSQLIFAVAQGDIDPEEALELIIRLK 497 TEEYRITIHNRNDSEEEILKKETKAITLIYRILRKAEERGLRAHTRLSRTHRNNTTTFRVTITVQ 498 PELVELLIEFVKRYDPELAEKLEENLKRLEEQGVPPEVIIFTLLLAVRAKDEELARRLFELVRRG 499 GEKYKEIVKSASRHAKEASKKNDKHLLHMLITFIKTIADTVPDEDVTKYAKKVATKLEQNG

[0382]

[0383] 500 DDEEVKTWEKLAKEAINRIPDKDTRTEVEKMVRMLIKTAHSPDPTEVALALSLVEQIIREFRGTEREAVKESLERAAKLFGVPSVEIDTRNGKVTVTVQGNDEAAKQLSFFAKFLALLAGVDLQVRHR DLQEEIKRALKEGDEELVEKLLRELSKRSGAPPALVLNNILVQLEEEGVIPPEELRKILEKFAKK DLEEQAQLTARFVAHLVGVESVRVTVRNGKVSVTVTGTDEKAQKASRMIKFALKSFGLDVKVQVR SEVSELARWVAANARAVKRRLEDPTKRKKAEKAEKLAKKAGHDPDEELARRLLTHLMMMLIEL SQIEKMLRFLTALARHKGDDKARKIVERAIERYRKGKIDPDQARKLAVKAFEILQS DEEEEKQKEVDRVFSHARRIPDKEKAQRFVKKYIEEARRRNDPKLVEMLAMVLTMIKALSEI SEELASDVLRLALEAGDMEAANRATMIAMALITGDLSDKEAKKLLKKLKKEISS DEEEKRVEELLRDPELIERFLAELLEEAVERGDPELNHLVLAVLFALRRGDKDLLKKLLEELIRK SLHELVALLWIRVPDPELEREILRVFFAIERGELDPKELARLIEELAKKVGDEEVLELVRKLLEK DVRNLLALARTLAEQAAREDSEHARKTIESLSRFVDKLLKNVRTEELKKLAKKVKKTIESLLRK DIVEEAHKLLSRAMSEAMENDDPDKLRRANELYFKLEEALKNNDPKTSQEIAKEWTWAQSD DMVEKLTTEAERVATKLERSPDPRNKHVARVIKSAIKQALSDSDKDSATLILFFAVNLARTF DEKKELRHLLKIVLELNRKSTDDEARTETQTLAFAAILAEEKGDLELAIRILRKAIKIAEESVK DSKEIEELRDKIRKIAKEVQNPRVALAALFVHTLLRHVRAGELDETRATELARAAFRQASQS SRLEEALRLLAEVAGLPPEEIRRIFEEYRKEAGDPRAAI FTLLLALRERGLI PPEVLREVIHLVE DEELELRALLIIYRILRALEKIHDEEERKRFMEDMIRKFREIFHVEEMIRLFERVLRRAAERAKG SEELKKRAEQLIRELKDKARKASRERERFILVALAASLERALKLNDEELLRRIIELIEAQERHQ DYRELMSEAVQLILKALSLPDPRAHQRAQRLAFILAHANNVPDETTAKQLIETAWREIQKVLG PEYEKIVETLTKQAEEAIRKNDEEKLSRVFELWRFYEEADESGDPRLRDLAHKLNHRVLRLEF EPEEKARTFEELVRKAYKKDPSKAIHLHNRLLWELLEEARRKNDEKLYRLAVELYVKLYRYLE TEIERKAHRLAAELFKKARKHNNEELAETALHLTTLLYALERARTEEERKRILEELERFLREWER GSVDELSKLVHKLYRIAFEKVQNPEERRKVQFVAFQAHLALKSSDPEEQKKAEELVKTLKKLV SLEEEVERLLKEAGVPPEVIELIKELIERLKERGFPPEAILFTIVFLLEELGNPEAARLIFELLR DENKDKLRKLKEKVDKASKNKDKDTLTKLWFEAFKLARDVGDPHVIRWAELAVRIFALIHR SEEEIKRAAKLLGVPPEELKRYLRRLLEDAGDPEAVKFTLALAIEKAGLPPEAEELLKKVLEILT PPDEYLKTLIRLVHELFTSGDPHKQHLASRVLTLVFQAHLAEDPNEKETLIRKAQKLIKKAG SEEEVKQRVTELIELWKKSGDDRFLEEAERILVKFYHEAAKRQDKRLIRTASRLFFKLEEIKRK SEEEAIEMAWSLNQLLSPEAREILKELLKRVKKGDPEATRIISTAFYLARRDPEEATEFIKKLR SKEEETTRLYELLLKNPEEFKKEIEELIRKTGDPRLEELLRLIEEALKEGDPFKLIFLVTLLLET SKEEEVRELLRLLEEGDPHAAMRLLFLVTEAAGGDPDLANEIYERLLRLLKKNEEEAVKLLVELR SENT RKAI EMI LNAAKNTARDPRIQKKIDKVLTKFKEAVKKNDEDKLREVAEQAFELSYQAVT SETDKVKKWDTLLTKAKQQQDPELAQEASSFAIEVLNKNPRDETVRELWKANMVFLQLHS DDSEVETAKKLVDEVKKHSNDPRVKFIATAVKLNLDEAKKNNDEERLKSAVEMARVLYKEAM DEELVEKAREALRKNDPRAAEVIFRLQIKAMEEGDEEKLRLAWALWLFAEGDPDKAEKVLSKVP SYEEMLKALLKEARKAGVPKAQKIIKKALKLLQEGKLDMDQLTRLMWEAWHLIS SSEDIKKTATKAYKKGDYEALFKASYIAWQEGDEDAASYVLRLLWEAAS TKELVRKLKELLEKGDPRWESLISKLMMLASELGDPELLHLLNLAYFLYEEGDPRVREVLKELEK DEEEIEELLKRAGLLSPKQVESLKEFVRRMKELGISPEEVLEQLLLTLRFIGDEEAVKLLESLLK SRRELFQRNVEKIAKSAGVPSVEVRYTNGKYHVHLHGDNRELKIAKNQVELFAMFANLDVKVTTH DRHQEFIRLIWETLRISRENNNEESHHLAAEAFLIWARALQNNIEEELKKAEEIAEKSLRLAR DAVEQAELFLRFMNSPVAREILKVLKELKERGVSPEEILRLLTEVAEKLGDPQLARNLRFFLEID SEHVLKNIKKWEHFTKEIPDSKQAREQAKLFIRFALRHIEDPDERRKVEEEATKLVQKI DEIYRELFRLAEELNDPELLMLLLTAAFMLSHGNPDVERLLKRVLEVLEKGDPEEAKKLLKKLLK DEIEEKLARLLWRALELGDDEAARLALEAQRLVARGEIDPSQAKKLAKKLEKLVRS DPRKKLKELVQRALRLAKEVGDEEILTLVFALHLAAERAIKNNDEDTLKRVEELLKRAIKKLES SEEEKKELEELVRRAEEALKRNDDERVKEVLQRLYELAARLEDREILHFVNRIHFKLYEKE DRELAHLLLEAAERNDPKLNQLVLVAFFLLERGVPPEEIERLLRELAEELGVPPELLERITRWR PLEEELEKALKEGDPDKLHRLLTRLMMEASRRGNKEELRLINIIYFAIEEGNPEMAEKAIKLLQK DELINEIIQLAVKDPEKAKELIKEIVKELRKKDPRLVKEFLEQLLLTLRFLGDPELVKLVEKLLR SPEEVLLELARKHNIEQAIQLLWLAAARDEELYKHVEELVRRNLEHVIVEQAKELLDRAR DKEAIENYIRLLKRETENLSDEELARQIRKKLKKAEELLRRGLEEEANELLLRIYFELRRR SFEELAKEYKRTGDPRYLALLFQLLFELQFETGDPRLTRLIWMLMEAMKRNDPDLVEKLARLAEK TELLERWRLVKEGDPEKAEELLARLYLRLAREGDPETLRFVIAVFLALARGNPDEAEKWRQLK S WEKAFRAAYLALNKGDEKAVREAQRALFLYYRGEADKNQLDKI I ET I I RRLRS DLEELVRELLERNPEFKRLLEELREAEEEGDEELVTLLRLTLALALAEFLNIPPEEARKALELLT SLDRALAAALERARELGDPRVNHIVLQIIFELRHGVNPEELERALVERLRQVNPEVARLVEELLK TTEEISKEIYRKAWKLAESVPDPHLREKAERVILTLSLMVLLIPNPEEAKKLAKKLEEIVRKIV

[0384]

[0385] SVEEILEELSKKLKDPRIEEVRERLEEIVRRLNVSPIIAAFILSFELFERGDPRASIWTALFLAPPSQEFFLRLVEKVVKKAKQIPDPRSQKIAKRLEELVKRARQVEDPKEKLHLVAVAAFLAAKVV SPEDEKLREELKRLYKIIKNVEDPKEATKIWFRAMEIFFNVTDPTLQTLARLVNHLALRVFLR SREEAIQKSVEHALKFLGVPSFSVSISNNTLKVTVKGDDSAARMAEFSVKSIAMLAGVPVEVRLK DKEELIEEAIEKLAPSPEAERYLREALKEFKELGNPEAFLFLLWFAAKRLGNPEVERLVRELLRR SRDELLRRNVEFLARVAGIPSVSVTVENGTVKVKVKGNDERAQFFSEQVKLFARFIGIPVQVDLR DRTEAARKAIDRAAKWFGVPSWSVDVRNGTLHVKIDGTDYSVKQLSFFAKFLAQLVGLPLKVDVR PEEEEIETRARKKATHWEVAKKNPEKAKRLIARLLVEAMEKNDHILNHWNHAYFLI EEVERS DRTETARRSIDRAAKWFGVPSWSVDVRNGTLHVKIDGTDDSVKELSLFARFLAHLVGLPLKVDVR DEHKELKELYDHILKLLRNAKTEEEVQELARKSVQLAELLNIEELVQESLLLWLRLHNRLK SIEEEILRWIMLAHKLGDPTLQSLAIEASHAYWNGDKKKARELIKKARKWY SEEYRKKLREAVELGDPELVAQLLLTLAFLTGDPRLQELFSMLSELIFRGDPSLEEILKLVKELG DEEEEKEKEVHKVLQTALRIPDKEKAQRFVKKYIEEARRRNDPKLEHLLRYVLMVIKKFSDT SFLETLKHLAEYLGDPELRKLLERLEELLKRGVSPEELLRIALRVARELGNPMFALTVEYLLHVP SDEKVKEALKEVEKAVKKAGLDPKWEWAERAAKEMKRRGISDDQISLFLRFWELIRKNSS SEELVERAKRLAGGDPRRALYLLLLALARRLNLPPRELAKILQKLRREAGGDPELLLKLLAELVE NYERVIAHAARLAAELGDREALRRLNEIFFGAREGRIPPELAERLARRIEKKLRS SLLSKIQELLERGDPEKVEELLRRLVELLRRDPDNEHLAMLLMQAVTIVAGGDPERANELFKRLR DEDEQVRLTVLYVAHLAGVPSVEVDRSNGRFTVTFDGDDDSARTASKTVKRMLTKMGLEVDVKTR DPKEEAKKVTRLLLKLAVNADDDEIRVELVTLAYALKRALERNDEEALKRVISKARKFIKDVKS SKEEWFSLAVKVSARYAGVPDVKVRVSNGRFEVTINGDTDSARLAQQMVRDLAEIMGVEVDVQVR SEEEIREIIETWKKDPKKAHRLVNRLLFEAYERGDPEKQSVAIRAAYLLYEGNPEEAEEVLKEIQ DLEELLEEALREAGFPPESIERAKRRVEELLRRDPREAAKFVALFLQFVGNPELLKKVREIFEKT PEEADKILEHLQKLFREALENRDEEAIAFIATLYLAAARAKRNNDQEALERVKKLLKEFKERKK GEEEALKILAEVLGRPPEEIRKLVEKIARELRVPPEHAIKHLALTIAFRLNDPRVLRLIAEALEK SEEERIKTSIERAAKWFGVPSFDIQTRNNRIHVTIDGDDDSARQLKLFIRFLAEVNGVQVHVTIK SLVEAVIEQVEKRGFPPEVLVATVWLALEKAGDPEKARLVQELAREVFRGELPPEELKRLLREIE SDDMRVLALAVSLMMFAQKQNDEKVEKTAITLWVQLMDARLRNDEETTDKISKLIKDLATKLKS DVEQLLLRYAWRALELGLEEHAVFIFRLLALVAKGEIDEREAHKLLEESRKRLEN SEEAIRMLQSLFKTDPEVQAQFLEVLIEHRVPDPEERRRLKETLKEIVERNDEELFERLVEEVLR PKEEEIKKLLKKLIKLAKELSNTPEARRMVKALERTVKELVKKTSPEDALKFAALALVSIEYIS NSEEIKKEIKKAEKKGDSQKLLQLLWLAIELGDEEAAVFALEAQSRVMS DEELKKQAEKLARLLVRLAAEVPNEEIALELITLAYALERASRNDDDERLKKVIDTAKEWSKG REEKIERLVREAERRIREGKISAQEVFEVQHRVYELLRKKGDPRVEKLLHKIMRLLWMLAK DEIERKLLKLSIIAWELGDQKALNLAASALFAWAGEIDPSQAKKLAKKLEKLVRS DKEELIEEAIEKLAPSPEAERYLREELKEFKELGNPEAFLFLLWLAAERLGNPEVKELVRRLLRR SEQDWRKNAERILKLVGVPHYEVSKSNGTVTVTVKGDDERARLALNQVKLFAMFLGVNVQVRIK PAEEILRELLHRDPRDKVWLLLYALLRAREEGRFSPEELIELLVRLARRAGNPELAKLLERLLR NEERRRAIERILTRAKKLARDPRIQKKIDKVLTKFKEAVKKNDEEKLEEVLILATTLWLRAHK KDELTEHVLKQLRQVLEEIRDYDRVREAAKLLAEFFRELDPRAEEAAKKALKIFEEELRKKQG SLEDINRLIFIAHKLGDDKALRVLSHALMAWEKGDEELARKLVKKARKLLS TEEIKRAVKHSVKDMVKNGLDPEWAVRVIRTFLEEMIRQGKISKEEAKEATKVAEKVAKKAKG NFFHWWRAQRIAMKKGDEKALSILARIAMEAAQGKLDPTKAERLARRIEKKLRS DHEEKAVRLLWAILEVAEKKDDEKLHHLVAQLYEQLVEAAERNDEDSLKEITKKIEELLKKASS DDHKEVIQSMLKSALRELKQYARDDSIREKVERLATEASRKNDHHLANRVLFLIVKLIKDS STVSREVLTIHTSNPEEVKKWKKEAQKRAEKIAKKYRKQGLDVKVHTFEVRRGDTVIIIVEVRIN IEEAKKLAMLVSMAAQMLAEGRLDERKWKKIVKEASKILKKLGISEEEIRNFQKSAKELKKQF SEKEEFIIENVKKLGDPNQIELFIAFAVKLSGDPRLKELLEIVKELIERGVSPEEILRIIEEFLK SDRVAQFLLFLAFQTGDPRLRILFSVWRLEEEGVSPEEIKEILEKVAKELGNEEVIREIKKLLD SEIANEIMRVLWEALERGDEELALKALHVSHLLWTGEIPDEEAKKLAEKLKKLLS DEREHARQRI EFLSALAQQI RRDEALEKAI RLMEELARWAGI EDI ERLVEEAI RRS RRAVKRF SEAERLLSHLARIAAEKGNEKALHWNRAIIALIEGLADEKQIRKLLKKAKSLLS DKEEKITEFSQKLKEKVEKASEEEAERLIAKAYHEAMQRNDPDLALVILRVMFEFREKRS DEEEAERLLAKLLRKARSPEERRLLTAVFFALREGDPETARRIVEELLRRVGDPELKRVLELIEK SSSQRYQIRVDSSDPKDTEQAHRDAKTIAQRYAEELRRQGNPVSYDLNYHRLNNLWMVWHVRTK KEWRDKLKSRLETWKKAQKDPSYEEEAERVI SEAMS IAMEEHDPEALHEANKAYFELHEYM PEEDEEFLKAAFRTAQEALKEGDKRRAEFIIAAAELYARHLDVEELKELAKELSEQFKEEVKR SEEQRKLAEKLLSKAQEALKKNDTETLEKWRRVLEIALKYDENPYIHKVAIDASLLWLMAAER

[0386]

[0387] SRDEQFKKLVKDVKTLAKRQNDEEVLKAVERATKLFREAKKKNDEEKALRAWFALAIALEKLSDELREILREALEKGDPRLVLQLLALLAFLAHRTGDPRLVKVYFIVAEALDKKDPELLKKALKLLK SPEKVLLELARKHNIHQAIQLLWLAAAKDEELYKHVEELVRRNLEHVIVEQAKELLDRAR SEHEAATESIKQAAKTIGVPHVEVDTRNGTLKVRIKGDDDAARMLQMSVKSVARLANLKVDVRTE DVEKLVKEVEKLARKRGDRRAAFIARAILAAFRNGKLSSEAAERALKKLAKRLKG EELKKILTELKKKATKALRNNDQSKVQHVIMVLAEIIHKAQTPEVRRTAAKLAFTLMRLTIQ DFTIHVQSEARWTVEEERRRALRWFARVSTRARRVADRLNVDVEFHVEHRFHGEKWIRIWHFR SEDDEKVSEIFLRAYEIALKKNDPDQLSELNRWAFLFVEAQQRNDEELVKRLAKKFEEWVSKAQ D E EAK YVH LMVF S L RY YAKQAKD P T LAKQ I KE LAD KAE RDN D P ET VREAL K WT KAQ KK S H DYQKVNKLLDKATEIARKQNNPDWFILVALKLSIEWAVKENDEEALKQLEHIASRFAKKF PELDRMVHMAQRIADKIVKKASNPEIQKKAKTVKKLLEEARQKKDDETVIRLLMIAELLFLSVF DDLKEKAKTLLELVKEASKKNDQKTLRELSLELVAVFMEAQDPNVRHLVEKAYFIAMELAFK SKEFVDSLLKHVEKLAKEVPNPEAKKILTKIKRLLEEANRRNDEAKAYLLVFTAWHQIRQLK SFMSVIQRAINLAMKNGDEKAMNILLRIQFLALQGKMSEEQAEKRARRIEKKLKS SEEEHVRKVIETMAKRFGVPDVKVSVRNGTVHVKVRGTDDAARQARLTAMFFAHLLGVPVSVTIR DEKVAVKVLWAFRLWDKTGDDSARKAILYASRVFMAAMNGDLSEKDRKKLEKTLRKIEKKLRS STIEFAISVAKFYLEELRKLGLISDEEAKRIKEELDELLRRGDERRVSQILNTLVELWRG SEEKAKELVEALKRRVEEAGDEELKKRIEKLVQEARKKGLSWEEIATLVFTLILAFETKKK DAVERLERQVEKLVKEVTNSGNPNSERAQRLLKLLKELLKKATSKEEQALAALTIALAVANA SLQEQLERIIKTIAKRAGVPQVKVRLRNNKVEVTVDGANQAAALVHLAWRVARRLGLDVHVRVK TTQETLVKSMIKQLKDVADNRDDPTVKKTLKTLAKILEKALKNNDMSAINRALFLAMEVLKSG SSKELKKRIEKAEKTGDVHKVMELALIAFMRGDQEAARLANRALFRLYNR SQDRREDAKEIIKELVKIGNEIDARSVLMFLAYEISEEEGLDEDQASEVLQELHEVLERLL NWEELVKRAQTLIKRAQDTGDKSYVNEAWSVANVAAFLAVELNDPEVQHIVSKLLEQIITVADS SKEEWERAVRLLEEGNPFEASILIQMFLLRLGISPHEIVELINRFMELFKRDPEEAKRFLREVG PHVEELVKTLVSKAEEAIRKNDQETLSRLIETLFELQKKDPRFRDAAAVALLLIAELQESN DLREEFQKWKKAQEAAKNKDHEKAIRLVWRALEIARKIGDEELERQVARVLLQVHELLR SREEAVELYHIAVEKGNEEAARIAAAVYLGLARGELDEDEARKLLEKVKKLLS DEELKRAAKSMLDQAKKLADQISNDDLRKKAQSLVEKARKIDDPTVALTVAFTVRTIYEEVQS DYREALRLLMAILELSERRNDEESHHEAARAYNHLSRARERNDEEAIKEWEQLREQLERIEG PERLRELLEESLRLAEERGDEELVNRTILLAFAFEIAKRRGDEERLKHLARLIEELWHENRN SEEDKIQTLIDEVRKAAKRLYPDDEEVKKIESNAQELVDQAQDQELAKLSLRFLKHLLDSE DISQELSKIVKSLIDELKKSPDPKDKHAAELAESAWRQAEEAQDPHERALILFFAISWRNT SRDELLRRNVEHLARLAGIPSVSVTVENGTVKVKVKGNDERAQFFSDQVKLFARFAGIPVQVDLR DDDKKEARKEAALFAQAVRQGRLDKRAAKEAVRRRI KKARKS GDERLAKI LELVLKLLDKI LRS DEILHKLARLLWRALELGDEKALKLALEAQKAVARGEIDPSQAKKLAKKLEKLVRS DEEERFREHLRRQLHELLRTDLRAAWIFVAYALERAKENNDELARKIAEELMREIEELIKN GPEEFIELYRRWLKLGLSPLTIANVILFAAYERNDPDLIRAAAIFMLLIQRFDPEEALRLLQELG DIEEFIETFLRSHNEDLRRLGTWSEEELKELTKLLRKLAEELDDEREIKNLLILIAFRSQLK DEQELVKKVKTILTEASKKNDKEKVREALSLATRVMLHAKDAKVFEQLFKLELYALHVIKS TRDELTKKNIEKLAKIAGVPSVKVTRSNGRWKVTFSGDDEAARDASNQILLFARFANLEVELTLK NLERKLVRLYNEAVKLGDKDLMSRAVAVFFLIEDGRISDDDIRKLVEEVERLVR DEEAFKIIFKLTVTVNKLLRKNPEQAKEFVKRVLKEAKEKNDHRTYMAAKAILRLIKEHG PEDDEVFHTLVRVLWKAAEDGNSEVFEKAFRLQFLFMEAKKKNDEETLKKVITKAKKVIKKQ SEEEHRKKLETELRKIREALKREEEAKKTKKWLRRLFREAVEKRDEELARLAALLYLAFERLR ARRYTLTVRVPTGDPTETKKAEKRAKKFATDVSEKWKRNGYTVRFRVDVLRNGDRISVIVIVIIK DLKKLVSRWENLAEEIKKQASPDEQKKLDTLLEQLKKALKNDDESTASKLWMIISEIWVRLR DEILEEIEEALRRGDPDLVERLISKLMLLAAELGDPKLLHAANRLYFALKKSPELVKEILEEVRK SKEEKMKEAALRLTLIAQRLGVDRKYMHTAVRIFVKLVKNGVDLDQARRIAEKWLKKWKS TITFSNTIILNGNSERVQKWAEEWLEEWRRKLSELAKRLGLEIKHRIHRERDGKFIVIRIRSRIK DESKKILDHLKTLLEKAIKNNDTETLNHLVLHLFFLARELNDEEAEKVAVEAYEWHRLLR P I E E E VKE L I K I VE EAL KNNDRETVS KVL FRAL RLAVEVNN E E L I S RVN RLAF I AFT VT DEEIEKLLEEVIGDPRIAREVLELFKELRKRIDPKEVLMMIIVLLNRLGVSPEVINLVLKILLLP DWREYSKKLKKALKRAAKDPHDAEAYHEIFRILLALEEIVPDPRIAEKLAQEASKQAREEVR SEETKRIKKWAKKLAKKAGLDEEIIKQAQKNVEELARQGFSPEEIKTVIKQLIWLAKKR DEEVLLELLQRTGDPRVLFILFLLKLLKEKGIPPEEALKIVEELIKELGDPEAERLLEIVRRRLP DPTHILLEISLLAQELLRRGIDPRKAVKIVKKILSKLIKEGKISPDEAKEALKMAKDVAKLLS DSEKLKEKVKKALKKGDVQALFEWELAMELGDQELANLATKALMILAKR

[0388]

[0389] DVRKLLKEIIEIARKQGDERAVFIAAALLAATEQGFSSEENSREAAERLFERLRNDELEKKVEKLAREAMQAVDNGDKDKVNTLVTELKKWKSQPDNPAAEFAALAITMLEYLIKH IEEAKKLKDKVRKAAEKLSQGRLDERKWMKIVTEAMKILAKLGISEEEIMHFSMQAYELKKKA SKKEEAKEAIKALRRMAKWLGVDEKYIRTASEIFKKLVKNGVDLEEALNFASIWLMQVLES SEETKKLQKKVREILKKAKSGKIDPHKALSLVMEILAKAMEKGDKKVLSEANRAAFKLIELESS DVELMAETAIKIARKKGDEKAVRILEKIAKLFRKGLIDEHEALRRINHIYFRLMK DEVEEELKKIAEKLSEAIRKNDEDKVEHLKEALINAAFRASHKYDPTRVQRIYRKVMRLAEKLG SIVEELEELLRRGEVDPERLVARFWFLLVREGDPHKALKAISLFMALKRGEIPPDEAVELLREIE TIRVTITWRVDAHNEDENRHSRRLAEEVREEAEEIAHRLNRRLDLSITWRFGDKWTIVFFTIQ TREILNKWEIVLKLVKKGVDPEQAVKIIRKILSELIRQGKISKDEARMATLTARFVAHLAKG LVIIVFYNVDPRLVKKFAKLIGAERITLFRFGQSVILFVKDASPELVKKAKKIVKPNRVTVRH SVEEWEEAKRRGELPPEVLALAYLAIFRAKERGLPPEDVARYVEKTARELGDKDLEELAKKLKK DPKKLDELRKKIETFLRKATDPDARMAAYFALNAAEKALKDQDPLAERAARAAYRQAKKAQ DKDKKIKDIVKTLVSRAKKDPRQAEEALRIVALLYLRAQREGDEHAAKLLRKALRLIHEAR SEEERKEEAIRIFIRYALKRAKSEEEAKEALEAIWRRAEKKNEEETLKLLKEALKRIEEKEKR SAVEQAELFLRFMNTPVAREILKVLKELKERGVSPEEILRLLTEVAEKLGDPLLAENLRFLLEHD DRTHHATFTARTTDEREQEELRQRALLWLDRRFFEAKRLGLRSRFRLSLTWDGDDWRVEAHHEVR SKKEEAKDAIRALRRMAKRLGVDEKYIRTASEIFKKLVKNGVDLEEALNFASIWLLNVLNS DEEVEKLLRRAEKLLKEAKKKNDKEKLQEAARLVLKAARLTTKREDLQRATHLAWRIAKLLN SDNLWTDIFELVMRALDEGVISEEEAKRIMARLNRAWERGLTEEVKKIKKEVKKLIKKARKS DVEEALRLAIREARKQGDEKAVKKLEKLLKALRKGLTSEETALRRASEIMFRLVN DSLMIEALRNVIRKIAKEVQNPEVEKLAKKVDKLLKLVRKGELDEREANKRALHLFFLAVSS GERYSLTIHVDQRDERLWREAQRLALLWAFRFIYEARRRGLEVHHSNREDTRGNRATNHVELQID TEIRKELEKIRKEVEKLQKKGRIDERLVEMIKRMIKQAKKTGDPFALMFARSLLEELKKLSG TTRYSSFYTSVAASPKEQKEIEKKAEKLAQKFRKEAEKNGLKVTVRVRTHRRNGWIVWEVTVQ S L E E I VK F WKQ VAT FAGAE D VKVRI S N GKVT VT FN GT D E S ARLAS RVI KNMLWLAGVD VKVKL K DVEELLEELKKLLSPELRNLLKVLLSQVKRIPDPEKREKLLRLIKEAVERNDEQLLQRLMFLAVR DYEIIFKLTKLLHEDPRKFEELVERLLRELGVSPEEIEKILRKVREIIKRDPNKAVSFVFQLIQR DEKVARTVLEVASELAIRTGDQRAMEAARFAAHVIERARRGELSEKDRKKLEKTLRKIEKKLRS DEKLRKIANWLMKEAAKAGVSRELIMEASLKLIDAYVKGKIDPKKLAKKLLKELRKEVKK TEEIKEQVKRAVKAAVRFGVDPEAAVRAIRLLLRLLIEEGKISKEEAKEATKVAEKVAKKAKG SETDKVKKWDTLLTKAKQQQDPNLARQAFSFAVKVSEKDPNDETVAKLVFKASIVWLQLHK TRTFRWHWEFKGPDEETRKTIEQLERDAEKFAQEFAEKYNVRYNVWFWNTGDRIQWILVIFI GEHRAQRLLWLAVELGDQKVAVKILKVIQLWQEGKISDEDLERLAKEAEEELKK DESVHKLAQKLLRKWAEAARKKDKKKLEEVEREAKEIADRIDNDEIKQQLMFFHFYAQKLKKDG SDETKRLQKIAKKALKLLRQGKLDIEEAKRIVQKWREATELGDEEAVRVAATIALALERIEEG DSLEKKFRTIARRLLREAERRRDEELIQKILLLWLAFERALENNDEDALKDITKKLEETFRKLQK SAKELAKRLVKLAKEVAKKLSDEEKSRKIKELAKKLEKAVKNNDERTVAQVQARLAILVLRAS DEIARAAIEEILERAKKLARDPRIQKKIDKVLTKFKEAVKKNDEEKLAKVYHFASRLFAEAQS SKEMQKWTKVTSKAHKLGVSPEAIFEITLQIARLFDEGKLSPEQIEKWADKLAKKIAKKAS S S EKAKKKI DKAI RNGDAQTAARALNEAYFRGLI S PKEWYKLAERWKARG DEEEKVAKELRKVLKDAGATSFRVMILNGKIWLFRGTSPEVKRASKWTKYARSQGWKLQVNVL SEEEQSRYLEMFLSHMFGVESVRVSVHNGTIHITVKGQTDAARAAQQYAKSYARVLNTKVDVKVK LVIIVFAHVDPRLVKKFAKLIGAERIRVYRFGDSVILFVKDASPELVKKAKKIVKPNRVQWH DEENLKILIDQSGNPEDAVHQLSLFAKFKGISPEEIEKILEKVLRELGKVSPEEIEELKRLLKEK QKHITITINLRNPHKDEEKEARERLRKFAKKWEEARKRGQKARIHETTVEFNGTRSVSVLVIIR SMEQANWLFVAAELLKKTGDDSARKAIRYALTVLHRAYEGKLSEKDRKKAEKTLRKLEKKLRS DREELLRELNRLAFLWRVIKDPKYVKTAARLFVKAYELANKGDEESAKKVQKSIETIITRYKS SEEEKRKRAKKKVRKIARSLGLSEEIARTLEFIFDSLVFLGVSIEMALKVIERFAKKLAKG SSKELKKRIKKALKTGDVEKVIELSHIALQRGDEEAARLASRALFDLVNR TQQVQIYVSYDPNDEHDEERAKEILEDRVRKWKKHQKQGLEVRVRRRTLNFNGNRVLAVRIEIR TDEREEVQELQKRLMDAAQKKDQEKLKKWKEAKDIAKKVGDPMVENLLRYAITVAQKAVS DEARKALVFVSLALAEYGVPHVKVNIRNGKITIRIEGDSPEAQTASEIAKKLVKQLGLPVDVQVR LVIIVFENVDPRLVKKFAKLIGAERITVYRFGQIVLLFVKDASPELVKKAKKIVKPNRVRWH SKELEKVIKKVNKKAKKLGVSPEQIKRIEKEIREAYKRGEHSPEEIERLAEFWAEFAARLAE DERLEHLKRLSERLFREALERRDERAIRIASLTHLALAKARNADKEEREEALRLVEHLLRLAE DDIQKEARKVNHQLMEIFFRYPDNEKIVKLVT HAAS LAL EAFDKNDEEILKKAKKLLEKVKKIC DKLTKAAKELAKKVKSGKLSPEEAAMWLSMIATRLGIDREEIHKAIRKLLKLIKS

[0390]

[0391] SEEVERRVKKLIEKAKEAVKKNDEKKVKDVLAILVSLMMEAQTEEERRIVITAWFIVMQLHHEVIVLVTFTRNGKRTSVTFRVDDDPREVKKAEKLAKDLAKKLKDQGVPLVLRVRKGDQRKRIEFR SDETKRLQKIAKKALKLLRQGKLDIEEAKRIVQKWREAEELGDEEAVTVAASIYLALRKIEEG DLKKIARKLLSIAFELGNHKIFTLVQRIFLAIKDGDEKTAKKLIKKAQSLAS DEKEKVRRVQKYATKLARKAGVSEEAIKILRELVEALIRNGVDASQAKDIARRFIEMTRFL SKELIEEALRLLEKGDPRVEKVIVKLFNEAVRKNDRDLNIEVIRVFMLLRNGNPEEARKLLRKLR SLAEWRKNVKRIAKMVGVPGVSVDVRNGTVTVTFDGNDEHARRASEQVKLFARFLGVTVKVTFR SIEELVKHWKFVARVAGVPSVSVKIKDGKVSVTFKGTDEAAKEASMLVKLTAHLMGLDVDVKFK SEEEKRKRAKKKVRKIARSLGLSEELAETLEFTFESLVFLGVSIEMALKIIERFAKKLAKG DKTELVKRIVKFVAEVAGVTDFQVTVSNGKYHVRINGNDESAQNVRSLIELSAFLAGIDVKVTVE SEEERIEKSVKQAAKFAGVESVSVRVSNGTIEVHLQGDNDAARQVELFLRFVSHLSGIPVDVQIR TEEEFLKTSIKELAKLVGVPSVRVRVSNGTVQVEVDGNDRAAKDFSQQLKLFARFAGIPLDVKIK SEEVAKMIKKNVEKIAKMLGLSEKITRTAKKWEKAYRKGVDPLEVTNLALQFLVKAAESK SEEELIREAIELVAPDPNLKETLKLTLRFAKHLGASPERIIFTLEKFLEDQGNPELAELFRKIVK DYRKAVQYLI RVLREALRKNNEELVQLVNRAMFI LVEVS KSNDEEKLDKVAS KLI KELQEAS S SSSTTWTYTHQPRDVQEAAEFLFKVLKEARERARRLRKQGIDVKLRVRREWRNGKVTIQVTVEEK NLALIAVFSLAKELYEKIGDPEIDKIRKKVEKLIRKYREGKLDESEAFNRAAKLQEELMKFI SEEETRKRAKKSVRHFARWFGLSEEIARKLEKLFEKLVKSGISAEEAMNFISVIAVQISEG SLLELLLRLLREGDPRVERVITRLLSLAIQRNDPKLLQWNLIYFAIKNNDPDMVERAIERAKEL SVTELVEEIVEKFKDDPEEVERLLQELAEKLKDPRLQQAVFTFKFALKLGASPEIIKETLRQFLK SWLRLAWEALEIAERLGDEKAHRAVARILHLLARGRISEEEAEKALKRLIKKLK SEEAIVRLYFKAARRGNRKVLRVLIRAVLALWRGDEETARKLLKLARKLD SEEEKKIRKLVEKLKRKGKLSDREIANYWKYWLKQGKLSEEQLLRATLIVQEWG TEEERHVEAVEVAARAAIQANDKEAIKKILEAAETVAKRISNPDLSKKVQKLIKKLRKALKS SEISEKLLKLLEKAYEKGDEKALQIVTKALMLLSKGEIDPDQARKLAEKAERILRS DIHSEYHIKIDPTDEQEEKRARLRAEALAYLAREKAEERGLENHNHIEIREHNGTIHVEIRINVH SQMEKARRAQKYAEKLARKAGVSDDQIEQLKQAARFLFAQGVDASQVKDILRKLIEKIKKS S EETKRI KKWAKKLAKKAGLDEWAI KLAQELVERLARNGI DPEVI KRMI ELT I RFMKKL DEEEAKFAVAWKQLAKELNDEETLKILEEAEREARERNDWRIVFKVAVKLYHLLSRLR DEIVELLFKLQYLALQLGDEKALRLAAKALMLLASGNIDPSQAKKLAKKLEKLVRS SVWIVDDERLAKLVSKLVNVERFQISTHGNLVLLNLVGVDPELAKKAVKTANKDAWRVH DVIEEILRLLRELGRLSPSVERALEFLREMLKRGVDPKDVLNQLLLFARFIGDPDVEKLAEEFLK DDRKKVEDTLRKKFKEAEKNNDEQALARWWKALELAEKFDDPELEKVAAELFETLARIQK SEIEKKLARLLVKAFEKGDRKANEIVIKALFAYFHGLVDPDQARKLAEKAERILRS DRDELIEENLRFFSEAANVPDVKFSVKDGRVKVTFNGDDEAAREAARAIRIFAWFAGVDVKVTTR SRDELLRRNVEKLARIAGIPSVSVTVENGTVKVKVKGNDERAQFFSNQVHLFARFSGIPVQVDLR TEQEMASHTIKFAAFLAGVSDVKVDVRNGTLKVTVRGNNESAQVLKKNVERIAKHFGVRVEVQTR DEEVKEILEELKKRGVDPEEVFKIAAKILEEKLNISPTEAIFIAQYVALELGDPELLTVAAKLIL DLWRLINRAAFLAVELGDQKVQQLALKAARAAMKGDEKTVKKLIKKAESLLS SEEDKIQTLIDEVRKAAKRLYPDDEEVKKIESNAQELVDQASDQEEAKLTLRFLKHLLDSE I EEVKKLMLLVS FAALRLANGKLDERKWKKI VKEAS KI LKKLGI S EEEI RDFQRI AKELKKQL DHLDEILKRIFELERKAWEIGDEELLRRLRLEQLTLLLRLAAHLDKEEREKLIKKWARLFQRAK TTVRVYNFEARTDDEELLRKARELAKKLLKESKEWAERKGGKVKNRISFDRRGDKIWIIAVSSVE SLKELLQEARERGDEDLLKVLSLLLFLIKLANIPPEEALKALERQLREHGDPDLLELLKELKELP EERVLSLLQAAIFVAAEKGDSHRAEQAMKLAILLFLAEHSGDEELVKKISTKAEKLAKKVLS DPLIFELLIRVQELADKFGLSPDEIKRFLNIAARLIVKGVTPEEIERYIKEELEKFKRKQY DEEKLERQARAALILAHQFDAEDEQARRFAKEAARKLFEAIERNNDELVEEIIREIERHLKESRG SKKEILEKVLELLKKNPDEAAQLILFLAFETGDPKVRRIFSVIAELIQTGDPELRKEAEKLLKRT SWEELYEKAKKLVEQAKKENDEDKLRKARRLANRAAFAAVELGDPTAQRSAFHLYEIVHSVAE DEHKAMLKTLIEELLRFAIELVDNPEERKKVTTLAKKIKKLLEKDPDTARRLALKLFREALKK DVEVLMRWEVANELLQRTGDDSANKAASWANRIIFRAIRGELSEKDRKKAEKEARKIEKKLRS DDILSEINRWFEAHRRGDDEILRKASRVAMLLLSGKIDDKEAKKLAEKLKKLLS DEHIRNTIDIHGTDKHARELVFRYIVESERKAREEAERRGLKVHSEWETRFRDGHVHVEHRVTLR SEEEIRRVIELIKKKNEMLAISLAFLLELVKRGEFSPEEFLNSAKRLVRWYGDPDAEELLKELEK DPTEWKRQVEMMAWLNAEFGVDPRQWKMVKKLLSEAQERGWSPDEAKEALKVAKKVAKRLS SRDETVERNVKFLARSAGVPSVDVKVSNGTVKVTIKGNDERAKFLTDQVRLFAMFAGLPFDVQVR LRSEVIKYTIKGDSEEARKHAEELIRRERHRARELAKRLHLRLRLFVKTTERGDTIHVDINILIR DKEQEDKVKEVIKRAKELVKKNPEEALSLVFKLWKEARDKNDERLMRLANVAAFEVHRYIS

[0392]

[0393] DIHKVTQLIFAIVEIAARKNDEELYKLAVKAYFEYFDARRKNDEKKSKEVTKKVEKLWKRAQSSVREVLFELIKIVRELLERGVTPEKIKRIVEEILRRLLKKGKLTPQAVRFLRKSVERALEYLEG SEMLATLVQNAAKKAGDKEATKIARKLVKLLREGKVSPSEVIEILFKLQIRLL FELIVFQLDFHKDDSSARKKFKDTVKKLKKKLEDIARKRNTTFSVHTHERRNGDIVILLVNFWR SKEDKAIRALFRLSEIAMKLGVDMKYIRTASEIFVKLVKNGVDLDQARRIAEKWLKKWKS DPNEEWLHQQAEMVRELAENLRDPRQIQDLVTAAEKMANAAEDPRVSKEVKKKLEEIYKKAV DMLEVLKAALLAYRLGLKEALREAQQAVFRLADGKASEEDADKELKKIRKKLNS DELLKKVKKRLEKLEETAKRQNNPEALKWQAVLRDVKEALKRNDRERVHILLLFAEHWFKRLS LLIIWFENVDPRLVKKFAKLIGAERIRLYRFGNSVLLFVKDASPELVKKAKKIVKPNRVTVKK SRAAAKVMKVAI KAFEAGDPKAVRLANRI LFALYRGEI DESQAKKLADEI LKKI G SEELKKLEKRLEEILRRLGVSPEQIKLLRKNLRKSYKISDSSAAKVFVEFVRQLFEKQRS PIKDEAKKIAKELITKAVKHQNPELAFEATLVLVRLAEQYGLSLEWVNEVLKELVDKAKKS PKWAEVNHILIRAILRAFEVGDEEVMRKAARLLFLLFEISDKNDEESLDKVEEKARKLLKKAI DETVSHLLLLLLEAAYLALERNDPEGASRAQRALFIAMHLAQDPDPKAQKKAQDIAKTVYSEVS DESEYETATRLAESAVKKVQDPEAEHLLRSIKKQWKELKSKYDDDEAVKYYHLMAYFLIQRFG DEYEMISNQLTLFARFLGVGSFKVSIDNGRIKVHVKGNDHAAEDITEAAKFFSKLFGIPIDVQTH DEEEEKEKEVRKVFRTAIRIPDKEKAQRFVKKYIEEARRRNDPKLEHLLRYVLMVIKMFSDT DIHHTVQIDSDKLGEEQKEQILRALRFTAWLALREFRERGIDVKVRITSETHGDRTHQRVHLRAT SSTESAVRLIRETVKILNPPSVTVTSSNGRVHVKIDGDTRLARQLQFFALMNSFMVGVPLDVQVR SVQDSVEFILRLLGVSPEIIERWRRIEELLKEGDREEVIRLIKKIAEELGDPQLELFARFLKAL SKKEKAKEAEKKLRKIAKKLGVDERMIREASRFFEINLMFGVDLDIARRMAEMYLKMMVKI SEIVKLLVRLARLAVELGKEKALHIVARALYALAKGAIDPDQARKLAEKAERILRS DLAS EALRLAFEAFKRGDRKAMHLALKAS I LI FEGEI S EKQAKKLI KRI KKLI KG S S DES VTRAVKFLATMAGVKS VKI RKRNGRI EVTVHATT SAAEDWNLI RLTAMRLNI ELDVKI K DETKKIQKEVKELADDARKSISDPRLRELVALARTLAQVAARDPRFKEMVKTLKRIIKELKKQG DEIQSKLNRLAFLAFKLGDRKAQSLAIKASHLVMDGRIDPSQAKKLAKKLEKLVRS SEEYEIRVTFDPNDREAEQEARLKVLFAVYRLLREAEERGTQPEAEIREREDQDSITITIKVKFR DHLKELFEHLLRLAFEASKTASEEQRHLLLTLLFAIRRALENNDEERAEKLLRSLEESIKRIR DIEEEIRSLAKKLLEIIRKDPRHKEEAEKTLVRLLFEAMKKNDQKRAAAISMAFLFLHESQ SEEEKKKLEKLFKKLIKILQNAPNPELRELARALERLFREAKKNNDAQALSNLVLRAKLWEKG DEEEEIRELIEKAVPDPELRRLLKEQLKRLEEEGVPPEEILFTLIFAALRYGDPEVARLLFELAQ DVEKQAKKAIKLARKKGDEKAVRLLEKIAKLYRKGLIDEREARTLILTIALALLL DEEVKKLLTEATKLSKEARSKASDEMNRYFLENTYRYLISQVKNASEDQKRELAKMAYEVLKAG DRRRRFRIAAALLSAKLLARDPRIQKKIDKVLTKFEEAVKKNDEDKLKEVEEQARKLFKQARK SEELKKLEKRLEEILRRIGVDPRAIKRLRKFLREVYKDTTSEQAKLFVRQVRHLAEVHRS DLRQANRAAFEAYRQGNREAQRLAVQLTSGLQDGNTSEEEAKDALRKLKEALSS SSEKLKRLAKEALKKGDEKLLSRLMSIAMELGDKEALREINRAYFEVHR SDEKEKLKKWIDKLVKKFGLSPEQRRILELTLRFAWRLGASPDIIKEILRQAAEMMKKRG DEELSEKIEEALKRGDERLWQLVLRLATLAFEKGDEELARRLQWLFLLSRDPEKAVRLVLRLV DYERVISRALELAAEKGDHKALRELNEIFFRAHEGKLDPTKAERLARRIEKKLRS DELEKLHTEAERIATKLERSPDPENKTIAEAIKAALRQALSDSDKESAKMVAYFAVNAARMV DEREKERAEFIAAALSLLAASSDEREREEARRLAEAARRELSRKDSRLAEYFRHVFEESVRRLG SREEWRELLERFPDPREVLVLLYNILIRLGDPQLAALVAFLFHKLENGEIDPEEVRELVRRIVK SREELLTRNVEFLAEQAGVPSFSVRVSNGKVKVTIEGTDEAAKFAVNQVLLFAMFMGVDVDIQVR SDESKRHSFLTKLVALFMEARDPKFQHLLARAYDEFFRALDRNDKETLDKVIKKLDKIEKSY NLEYDALRLIFLAFLKGDEKARTIARKLQAAIMKGDEDAIKKLATKLKRIL SDETKRLQKIARKALKLLRQGKLDIEEALTIALRWYIARELGASEAAEEAFRIQYALLEIEHG PREEVLKILEELLKAPDPEKVIARLLLHASREGDEEKNRLVLAVIFLLREGVPPEEIREILRKLT SEVARRLLELLKQGNEEEAQHLWRALWEAAERGDSELALLLFRVQFLLMQGDPRVEEVLKRLEK SSSQRYQINVDSSDPKDTEQAHRDAKTIAQRYAEELRRQGNPVNYEVNYSELNNLWWFVTVHTK SEDEKIKKHVKTRIKQILSKNSPEARKEAANYANRWFHLYHVPDPHKQKLANEIFHYWSRLR LEREHAIKRIVEHLVKHFGLSRDIEKTLKKWEKLLKKGVDVFRITLIVSQLADEWARRLK SDESRRELEERLEEAVRRGDEEKLRLLASTLAYAFLRAGKLSSREALKYVEELIKELKKRV S KEEVLS RMARLT FRFAGATDVKVT I RNGKVTVEVRGDT P S VRAAS RMVTWLAKRQGVDLKI RVK SRETLKRRVKEAYKRGDAHELFKILFEAMERGDDEVIRFVLRVQFKLARR DHRIRITFHFTGDDEKTKKEIQKEIERIRKELREEAKRKGKDIRIFVQRRRHGSTLVIYVWRRK SEIMARAVIKLARKKGDEKAVKIAEKILKAAERGLYTHEALHKLNHLIFYLMRK SEEETIEEIEKLLERAVRELRVEELRRLVEELLKEARRINDLRFFEEAAQIALLAWLQDAL

[0394]

[0395] SELREINRLMFLAVELGDPKLAQIALKAYRALRSGDEKTLKKLLKIAHKLVKSNAERWRLVKKLREELNVPPEEAIKILLEEALRRGDEELVFYLIFLLILVLGDPEKVREIFERLR D LAREAS H VLAQ LAFSAEKTHNEELS RLQH RLAAL LAEAI IRNDEELLKHLI RLARE L L KRVHN PEDDEVQRTLQSVLFKATKDGNEEVIRKALKLNFLFFLAKRNNDEETLKKVITKAKKVIKKQ SEDDKAHKVLQTVLELLSKDPKKAQDFVKTFLKKAQKNNNERLVHLARYILMQAQMFR TELVELAIELAKRLGDEMLWSLMFILELVRRGEYSPEEAEHAWRLARQFGNPWEIIERLLKK TERELIEENVKFLAKLANLSDVKVSFSNGKVTVNINGTSEHADDLRRQLTLFARFANLRVDVQTK SEEEIRELIEKLGDPRIAI FTLLYALERKGKVPPEELHEALRRLLKLLKEGNEEEVRRLLTELID SEEELKRLVERWEGDPDARRILVELYNELVKKGDPQKAIILAFILTLAEEGNPEDAKRLVELLR TEERRWRWELRKPSRQEEDETRAEALRVAYKALREAQERGGDAHSRVRVKHDGDTVTVETRVEIR DRTETARRSIDRAAKFLGVPSWSVDVRNGTLHVKIDGTDDAVRQLSLFALFIAHLVGLPLKVDVR SDRAFRKAVHIVEAAARRLNLEWAEELAKHLLRLWEEALRRNDEEALKWLFEELSRVMYRLAQ DWRIAHRLLREANRIAYELGNERLWQLVARNFLALAKAIKNNDEELLKRLIESSKRLLEEIKR SSSTTWTYTHQPRDREEAFRFAAKVAKEARERARRLRKQGIDVKLRRRLEWHNGKLTIQVTVEEK DLVRLAVTAFLLGDPEAASLINVLIFALVEGDPEKLREALKRAEEVAKRLNDPVLEKAVKIAKKA PLEEELERLLKENPRAAVTTALFLAQERGDPDLFKELLELLRLLEEDPNDEEVIRRVEKILRELQ SLEQAIEFLLKTQGDPAARIVLEFVRELRERGVSPEEVLRLVEELLKKLNTPAARQLLLFIMFLK SSAHVIIEALKLARKKGDEKALRLLNRLAFRAVRGELDPTKAERLARRIEKKLRS DVEELVEEVLRKNDPRVLEKVLRLLREREGDPIRAFYILIELIAKILKVPPHEAAEILHRLLRLL SAEDDVTSLVRLSAWMAGVPHVEVSFKDGRVSVTLKGTSPSVRSAKRVIQRVAKMMGVRIEVKTR SIEEDVKMIVKLSAFLAGAEDVKVRISNGKVTVTFNGTDDAARSASRMIKHVLTMLGVDVKVKLK DLEEAVERVIKRFKDDPENVAILLAFLLLLALEKDPRLVEEALRLIEELLGDPELVKWLERAVKQ SETEILRVLFIAHRKGNREVAELALRASVAAFLGDPKTAEKLLRKAKKLLKS DEDKELEEIVKKLAEDPTVQEIILKVASNSPYLRRTIIKLFRFDPRQAHLFIRFLAHQLK SIDELLEKNLRKIGDPNLLRLTAVFIVSEALNVDPEEARKLLEKLERELGDPEEALRLAVRLALK NLIAEAVWKIARELYEKIGDPEIDKLRKYVEKLIRKYREGKLDEHELNERALRVLFKLYRFI SYEEILVKAYFILAEQFFERPDDEHIRKALLYVLRLVMKAIKASDPKESHEIARQVEEFLKKVS KDIEKEIVKVFHTAMERGDIELALKASLVFMALHRGEISDEQAKKLAEKLKKLLS DELLTEIIHLAARDPEKAKELIKEIVKELRKKDPRLVKEFLSMLLITLRFLGDPELVKLVEKLLR SRVEKILVRLIARTGDPKYTLIAFALILLRREGLPPEEILKIVEKIVREANDPDLLRILEEAIKR DIVEKVRRLATVAIFVAHKQNNEEIAVTLMKLIHQAHEAKLNNDEETLDKIRKIVETLYKRW SDETKRLQKIAKKALKLLRQGKLDWSEAIRIVLKVAKIAWELGDQKALNVANSIAFILVKIHKG PEESEKFKKTVETRVKRAVDNRDMDLVNRWWEALEIARRLNDPELEKWAAKLWLKAVRNI SEEFRRHIEFLNRLLHLAHHHRELAEEAIRLLREHAEHAKRLNNEVLAELSERLARQLEEAIRH DESERVRLAFWAAAHAYKHAQSEEEKKLAEAALRSAVKFAETKSDPKMQKLAKKLKEKLKKIV SEKEAKKLLKIARKIGDRRAAFIAAAVLASLRRGQLSEEAAKAALAWKELLS SKEELKREVAKLLGIPPEELEKLIELAERAAGGDPEIAELTLYLALARLGVPPEVIKRILELLKR SEEAAHLVQVAAKRAGDKEATKIARKLVKLLREGKISPEEVMRILNRLMFRLV DEDKKVSEKVKKLVRKAENANDPTQIFKLLRKALELVMQVSDPTIHKEVATEAQRALFIAARR TVRWEITVSFDRHNEEEERKARELLRRLLKKVHEWSRKRGLRNLYWRFETTRGDRVTIQVTSLSL SLEEIFKVSFIAWRKGDEEVLKLAVRAIFAWLNGDPKTAEKLLRKAKKLLKS TEQEKFKTTVKRLVDQALKENDPRLAVRVIWEALEEAEKKNDEDLARFVARLFLKVSRWFA PEEVAKMINKVAESQGDKKALKLARKILKLLREGKLSDEQAINELQRILFHLA DASDIFRLLEIAMKLGDEKALRVLLRAQFALAKGDEELARKLVKKARKLLS PEVEIALFFAESVLKQAKRDNDSRSVRWISILKRVIKKAQTEELRKKAKKLSEIAKKVEKS DIQVAFTVQEVALQLLDETGDDSARKALNYATEVLFRAVEGELSEKDRKKLEKTLRKIEKKLRS DDWTDHVEKLFSKAMEAVKKNDEDTVKTIEKILKDLTKKAQDPKQRELAALAITLVQSLLKD SLEEDLSRMARLTFRFAGATDVKVTIRNGKVTVEVRGDTPSVRAASRMVTHLAKHFGVDLKIRVK TSRNRFTVEVNKKNEEERRLAEENLWILAAFARRRAREEGGRAKVRVEEHEDGDTTRFHNEVEVR SEVEKILVRLFNIAMKKGNREANRWNHAYFAYKRGDIDEKQIRKLLKKAKSLLS TRVFTWRFQAKNVHEQEIREAREHLKKLADHWLERIKRRGLRARVRTSSFRRNNTHIIWQLEVH DEEHTKELFEEAKRIAEELNIEELKFFVAAALANQELARNIDDEEEARELTESAINILRLI NSDELKKRLEKIYRLVEEGKLDPDKAEQIANEIWREARKVGDDSVRQEALRVALYLHQFKK SKDELITRNITELAKMAGVPHVEVRVSNGRVHVKLKGDDRHAKMAKRLVELTARFAGVDVKVTIR DETLEKSLRKMLRLDPNMFELTVMFVLKLRGVSPEEIEKILRKVREIIKRDPEKAVEFVLRLIRK TETHHFIWTIDGTNLRAAAKLTTLAAKEARREIKRAKRRGISVTVTITVRVNNGLAHVELILRLH TEVEALSRIIEFVARVAGVPHVEVSWSNGKVTVTFKGTDHRVQEASRMALLTAMFLGVELKVQFR PTEEIQHLLVRILKKAAEDGDRRLLHAANHLYFELEEAKKRNDEEKQKKILKRVRKLLKEAQR

[0396]

[0397] SEIMELMTEAVDLGDPELLRLITLVAMAVAKGDPELLKRVEKEVRELSRKNPLVRRLLKIIEKLKSIQEEILRLQFIAFEQGDHKIARLALRAAEALLNGDEETARKLVKKAKKLV DEKKAAKFLKKHLTERARRAGVSPKAIKKIQTIFDRLIKSGVDLLEATLIALKLIDKWRRS DERWMNASLLILALLNRGVSEEQIKKFVKRLLEKLRKSGKISEEAAESVKRLAKRLIKIARG DLEKEAQRVLYLLMHLAMQIQDPKQRHKAQRIIWRLLEAVKKKDEDKIKKLVDLAKKLIKLV SEIVKKLLKLAEKAVRQGKRKALRIVSTALFQLVSGDIDPDQARKLAEKAERILRS TIEFENTIVLNGNSERVQKWAEEWLEEWRRKLSELAKRLGLEIKHRIHRHRQGKFIVIHIHSRIK PELDKIVHHALSLAAVIAFKATDPRIIRKAMKVSVLLMEAREEKDKDLVTKLLKKAQTLWKLVS SVSILLLTAIFVAHEKGDEKALKKLQKLLRALQRGLTSEETAKRRAETILKRLRS SLVEKIEKLLREGDPEEARQLALLLWLALHEKGDPRVELVRRLFRLLAKGDPEKAKKLAEELKRT SIMEEIYKIQRLAEKLGDPKLEQIAMRAALAAASGDEKTLKKLRKIARKLIKS SPQEAMELLFRAQYIARQKNDEELTHKLVRLLWLLVRAIKEGDKDTITKIERLAHEIYKKC SLTETIQTSVTQAAKFAGVDKFSVTVHNGSVKVRVRGTDDASNQLILFVRFLAHRANLPVEVEQE NELEKKAQRLAHLLAEIIKKVPDPTIQHQAATLFILLFRAIVKNDEEKLERLIKKAEKWIKSV HRHYQFSVSVRGTDERAKKILKKFKEEVKKLTEKARRDYNTKVTFRIRSLRFGDWLVFVDIRVE TEQKSEEIETDGTDEEAEKRAREFWERIRKQARREARNRGRKVFVFIFREEHNGHIRHSVYWFR DLSKELEQRVRKLEKKWQEGKIDTQEALTEVLTLMFKAEETGKLDPKYLEWLLRVFEEWLRRVY TFMDVINRAIFLAHKNGDDKALRILIRITHLAMQGKMSEEQAEKRARRIEKKLKS DTKKKSKELLEILERLAKRHNLEELKRNIEALRLALRAAQQHNDEELIRKVDEIAEEAIRRG SIRFKDYVQVSATDPHQVTRAKEIAKKFVKKVEKEARKRGLSVSYDVFVRRYGNRVFIVWIEVE PETVELSIRLKDTDEEAEKRVEKWLETIIKKLEEKAKKLGLRVKVRKFKLRRGSTLIIQVRIEIE DRKEVQREVKERVDTALSNNDPRTAAQWLDASRLAEELGDEELLRLVSQLMFKIAEFQG SSEELKRLLDEARRRGDADLAAEVAWKAAEEGDREAIEAAFRLQYELLN DDWTEHVKKLVKDAEEAVKKNDKHTWRIWLILFRLFIKAQDPKQRKLATRALALVLHLYKS GPEEAKEILKKINSPIAREVLRLLEEAERKNDPRLLEEAILVALTAAFAGIVSPEVFRLLVEFVE NEQKMIETLIRSAIRDGNERQFRELVKVAKKHGLLSEDTLEQLKKEFKRFKKY TETRKITITVPGNDEKAKKILKDIVTKIKKDAETHAKKRNLRVDIRYNTLEHNGKITVTIWIFK DREWHAHIDVKGTDRRAKEIAEHWALTWAFRILLRTRRQGLESHVSIKVRRDGDTVRARVTVRAH DVRTLATRVKQRLTELYKLDTPEARKEVIKVWFKLMEIYSRITTEEEAKIILEAVLVAISVLLG SDLARLMKLSQRAFEAGVPKAERIILRAMFALHEGKLDEKQLRKLQKLAEKLIS SPDEIATLVRKNVEKLRRTFDNDWKRQAKLTVRFAILLAKKNNDERLVRLLKTILKQIEKLSQS TELVEITLMFASRHNPLAKELLELLRELKEEGKFSEEELIKIIVKRAEELGNPLLARNLEHLFRV SSEEIKHLIALVAFIFHKKGDRRAVEVWNRLFRLLELVRRGTLDPDEQKRLLKELKEEVKKL SERYTFEVTFDKNDKEAEERARQKLLLFWARVAFRAERLNRSAEVRVRTSEHNGTIRIHVTIEIK SEELLREWERLGVPPEVLEVLTKQLRRLEEDGIPPEVILFTLALAAERLGDPELAELLAHLAAR SEEIEELIRELFKKARNPEVKRVLALLYLALKKARSPEEREEIERLLRLIEEDPRLAREILKKLD SWKDLLAEINSAFMKGDEKKVKKLKKIAEKRGDERAAHMAESALRQLQRR DELINEIIQLAVKDPEKAKELIKEIVKELRKKDPRLVKLFLETLLLTLRFLGDPELVKLVEKLLR LDEEKRIKTTVERLVKHFGLSRDIEKTLKKWEKLLKKGVDSEEIALMVSMLAQRWAERLH SEIEKKLRKLKKKARELGNKEAALAVQQAIQAYREGRISPEAAKRLAEQAERILRS DEEVEKLLRRAEKLLKEAKKKNDKEKLREAAHLVLRAAALTQSDEDLKRAARLAMRIARLLN DEHELQRLAWLALELGDEEIAREAFRVIMKVIEGKLSEKQAEKWAKKIRKKLSS DSEHMVELTARFLAKRVGVPDVKIRKSNGRITVTVKGNDESAQIFSRVLKHMAKRLGVDVRVQTR DKKYNELLRLLWKALELGDEKAARIAFRALVKLWEGKISDDDYKKVQKLITKALRE DEEVEKLLRRAEKLLKEARKKNDKEKLKEAAKLVRKALRLTTSEEDRIRATRLWLRIAELLK TLDDVMLRAVESQARKLIKKSRDPNAQSKLKKAIKIIQKASQVSDPNIKKSLVQKALTWMDAA SEEVKKILKRIVKLLAEQLGLDEKAQKIAKKVAEKLADKGVDPLEATLILVRVLDTILRRKK DEELKTLMNIVNRLMFEAWRNNDEKSIRLAWTALQLLMRAHNADEEQQKKLKTLVKQLIKKIK SEVQELVEEVLKRGNLEELIKEVLKLARELGDPYLEAAARDALELVKRDPERASFILVLVLEELK DKEEIRQKIMNLAFAANLRVQDPRIKKTIHTVYRQAMEALDKDDEDTLRTLLKIVEKLLRDVHS TEHYRWTVRVEGTDREAHQIIDEFIARILNEAFRIALKDTSHIRITVHTKVKDGEIRVEVRAERR SINFDVQVRDENNHFSVLVQVEDPTEAEQQVLLFLMFLGVTATVRVDPDTGSVEVQQ DDEELQRFIQRHVEETVGAPDVQVSVSNGTVKVTIDGNNDAAQAVRFTAELAKQKAGRNVDVTTR HEEIANKLSKKVAEILDQQNASDEQKSKAHEILLEASLVLTNISDKDKAKSYADKVTKDISNM D RLAYALT L S RS L L RDAKARNN E E L I RLAE ET S RL I E DARRRRD E E LARRL VH KL L EAI RKA TLREEAMKLAMEAMKRGDEKALRKAQRALFLMARGDISEKQAKKIIKEIKKLIKG SETYKVTIDFDGKDEDTRERAELFARYFRLRAEEEARKRNVEVRIEIRFDTRNDKLKTDVRAEFR DEEVLELVEELLKQGDEEEVEKLLAHVAARLKEKGDPEKAILVTNVWLRLKKNRERALEALKRVP

[0398]

[0399] DRETEIFEFFAKTTIKRAVEENDEERLKELEHAIRRFYKEAKKRNDEDKLKVAKRLLEWFKKFV965 DLEEELEEALKRGDPELLKRIVEKIIKKLGLPPNAEYSLKWWLEFLLRTKDPRAREAVESQVRHG 966 IDDVRWAASNARAYWARLGDDTFDKLRKKLEKVEKKVKEGKLDPEQARKFALEVFRRLVKLV 967 WI I FVDN E E LAKQ VE E VARRL GVE VRVS KQ GD L VWT LQ VRD P T LAKT AAS WP EARVH E F 968 DEEKKELHKEIERLLKELLRHAKSEVEHLAAASLLQLHRKAWENNDDELWEESKEKLEDARHALR 969 SVWWEDERLAKLVSKLVNVQRFSIFTFGNWILNLQGVDPELAKKAVKTANKDASVLVF 970 QEEVHRTLLNLFLMLSHNARDPNLQEKAQRLARKALQAVDKNDDDTIKTLLKLLKRLVDEVRS 971 SEKDVKRFIKKLLDEARKKGLISEALKESLKRVAKELLKRGISTEEIKRILETTARFLAKENK 972 KDIEKEIKKVITTARRRGDELLVAQALAVAHLLREGLITDEEAKRLAEELKELLS

[0400] 973 SEKDVKRFIKKLLDEARKKGLISESLKRQLKHLAEEALKRGADTSEIKKLLKIAAKVTAEFLK 974 SEEERKRLLEALERGEVSPYELALRLQRLAYENNDPELLNRATKWMMLAHGDPRAEELLKELLK 975 SLEEWEEAAELLNDPALRELVELLKEARENNDPELLTILVTNVWLRLHELGNPELARRVAKLRP 976 DEEESAQRLVKRTAEMAGVPRFDVKVSNNTLTLDVTGDDENVRDVTMNAHTTAHLVGLEFRVRTK 977 SIEEQARRWEHVARRYNVKDFRVSVSNGKVSVTFKGDDEAAKRASRLVELTAHLAGIDLKVTVR 978 IELEEKVEHLASRAMSVAMENNNREELRRVNELYFKAEELSSSPDPKESKEALDRVRKELEKIV 979 SREEELLELWRFKDDPKVILHLLEELLRLAQEKGDPDLEQLVAELFAILARDEERWELAEKLR 980 DESLKKFKSDVEKKAREAKSNNDEKRLSEIVEEVLKFAQEAQNNNDQESASVAAVALFLVFKIQ 981 DEVLEASYLNKQLLELARRLNLEELVRLAEKSLRDLARAALNNASEEFKKTLVELSKELEELIK 982 DDEEAEEAKKLVDKVSKTNPRFREQAQLVKKNLTRAQRNKDKDDVEAAILLIISNYNVAS 983 DKREEVKTLYKILEEAAKKRDEETIDKVAARAYHLAVEIGDRELLLKIVSLWMHAQELLS 984 NEAQEKAAEIARTLAEKARRLGRPVDVSVRFKNQTLTVKVGPDPKEAEEAAIRIFAALFAKLEG 985 DLRFTFSVRDDSDDERSRRQSEDLAKAFVFTAALAAIELGLELRFEIRRLRDGHSNEVRVEVRIT 986 SFAEMVLEYLEKAGNPEAAKAVRVLLELLKRTGDPRIRNLINQIAWAVMEGDPRREELIRRVLEP 987 SEFVIITVELRNVSPEQSKKAEKKLKDIAKKLTKTAEKRNLEVRFRIKEQSHRGTKVLIFNLWR 988 SEIEEVLELIRRYDPRWEMLLVSLMFLKHMGASPEEILKYLLRLAKILGSPLATELLRRLLEKR 989 SEVRALTLALQVELLAKKVGDEEAKEAAKKAEELFRRLREGKLDASQVEKLLKRIIKKLRKLG 990 RRHWEYRIRVDARDERQRREAKALASIIATRLWLRSKEQGQENHFHIREERHGEDLTFHIRNETR 991 SKDELIRRNVEFLARVAGVESVSVDVRNGRVSVTLKGDNEAAKDVKNQLELFAMFANLDVKVNIR 992 S KELEKVI KKVNKKAKKLGVS PTAI KDI ERFI REMYKS GLAS PEEI EKEADRLAKLSAFLAR 993 DQLEEAIRRLLELVPDPRDWNQIELFLRFMNSPEAREALRLLEELRERGVSPEEIVEIVLKIAR 994 HLHLRFSFHSEGDSERSERAAEQFAEAVRFTAALAARRRNLEVRVEIRIHIENGRTHVEIHVNIS 995 SETENVTFEIDGTDERAEEQIRRIVEKERRRAEKHAKKRNLRPHVHVLTIRINGKLLLIIVIVFR 996 SEEAKRELEEAKKRGNLERHLRSLLFALAAFLGDPEEATKRLEEIARSLNDPEAEKLVKKLSKET 997 TKEEKLKKAKKIVTTLAKLANVDPRQIKLLQSLVKMAWEKDDPFAKTMVEVIIRQLKHMVDE 998 DMRERSVEEMLRIIETVARRRDDPTAKTLVKRAKKILEEARRKNDPKLLRKALRLANRAYFSLK 999 VEELKKAQKKLDKAIKILRKKNDEMAVKLAEAVKRELKKASSDSDPRTIHRVIFVAVNAVRNAQ 1000 TSQREDRNEFQPENEEAERLFKRAIRQAARAAVQRIRQQGIDVKVRFRTETHNGRVTLSVTFRHR 1001 DEKDDFSRLVKLTLWLAGAKSVDVKVKDGTLDVKVEATDPSVEFASKTLTKWAKDQGLRVSIKID 1002 VE E L KKAMKT VQ RAWT I LMD KN D I EAAL EAVS VFMMLAKAS SDSDPRTIKTVID LAKQ VYKKAT 1003 TNEEWEHLARFNPRNLELTLRFMLELARREGNEDLVRVLEIVLRLLEEGVDPEEWRVFKELLK 1004 SDETKRLQKIAKKALKLLRQGKLDTHEALSIVMEVIRIALELGDRKAVQVANSIAFLLIKIEKG 1005 DLRKVTRLIWAIAEIAKRKNDEELHKLAARAYAEYAKARRNNDEKKSKEVTKKVEKLWKRAQS 1006 SKELEKVIKKVNKKAKKLGVSPESIKAIERNIRTLYKLGLLDPEEIEFFADMQAHMAAKRAS 1007 DEEDKKFAEKVRRKALELIHKDTDEETRIHLTNLWLRAEAAQKTTDSEARRLFKELVEELERRL 1008 PEVEELLVMAEIMLKQAKKKNNHMLVRSVLRILKEVIKKAQTEELRKKAKKLSEIAKKVEKS 1009 RIEIEHSVLIRGDSEEAKKRAEEHQERFRKFAEDLARKLNRDVEITFFITELNGVIHSIATARIE 1010 DETHKLYITVLSLARQAMSKNPEYHEEVARLYVKIFRLWFKGSPRVNEVLKKAEKILRSIIG 1011 DSKTIAMLRVLIRIYAKRKQIPEIEKLAKKVDKLLKLVRKGLLDETEAEKRAARLWFLAVSY 1012 HLAEMIKHLAKREITKAVQNNDEHKVELWFFLKNAARSVDDPRLEEEIKRLVQTVLEQVL 1013 DPDEILDKVANTLLTLAFQHPDDPELHKAFSMVAEISQKAKKLSDPTQKERLARLAQRILEEVS 1014 DEEVAMTVSKVASMLFLQTGDESAVKAMHFALQVITRAEKGKLSEKDRKKLEKTLRKIEKKLRS 1015 REQFFFFVRVNGNDPTQKKKAEDKAEKFRQKANTTAKRKGKKVRVTHSSSTQDGKYWLWIRVE 1016 SEFIRELLRVAKELGDSVALMFLEQLLELLERGELSPEDVLRLVLLTLRFIGNEELVRLAEEVLK 1017 DLDKRAKIVLSLARHLRKKVQDPHIRKLLEHLAKILEEAVKKKDEELIKKILVLVFRVIVEAQE 1018 DEEEKFRKTIEKIAKKLGVEQVFVKVFQGQFWFIVGDTESARKVQKLAKEVAKKLGVRVRVRIF 1019 DKELKKVLNKVMKKASKLGVSPEAILHIAIQIQRLFREGKLSPEQIEKWADKLAKKIAKKAS 1020 DMHEEAIKMLLRLLETARKEAPDPEKSKKIDKIIKKAKTWSSQVSLEESANYIFDLWKVQRE 1021 DEEERRKISERLKVDPKVAESLLRIKKWLERVLDPEFVRHSLLMFIFLLLKVDPREAERLLKLLE

[0401]

[0402] 1022 SSTQRSEITVQARTEEEAEEAEEELRHSAEALAYLARRRGVDVKLEIRIERRGKRIRIRIRFELR1023 SSEELKKKIKKALKDGDRRAVTEIAMLAQELGDRELARIALEAAEKLAN

[0403] 1024 DPTSKKLKKEIKKVLKLFKKGKIDPVEAWNLLFRLIEKARRIGDEEAVQLAMHAIAIILHAMKS 1025 DKDESFRRIVEWIARMLNVPDVKVSVNNGKFHVHLKGNDENAKDAERLVRLTALFMGVDVKIDLQ 1026 PREEILHKINHLYFKAHEDGDKKVLKLLAKLLYKLMDAVRRNDEEKQKKIAKEVEKLEKIVQR 1027 DERRTFTLEVKGTDEKAKEKLRRFALEVQFRVTTEARKNNRTVRVHVRVERRGDKIRVKIDVTIR 1028 DPSEEVTKTVQSILTEALKNKDKELVHKAIRIALEAARKLNDPRIQEVITEAFTVALAVQLG 1029 SEEELREALKELAERGDLHAAARVLNSLLFLLYEKNDPSMRKVAEVMIRANEDPRRLPELIKLVH 1030 DEKLRKIMNWLMKEAAKAGVSRELIMRASMKLMMAYEKGKIDPKKLAKKLLKELRKEVKK

[0404]

[0405] 1031 GDRRKEMDKVYRTAYKRITSTPDKEKRKEWKEATEQLRRIAKDEEEKKKAAYMISFLKTLG

[0406] Table 8. GP130 binders DLSERIRRLIRRADELARRGNPEEARKVLEEAEELAERSGSPELLESVRFLLEVLG ( SEQ ID NO: 1032 ) DISERFRRLMRRADELARRGNPEEARKVLEEAEELMERYGSPELLESVRMLLEVLG ( SEQ ID NO: 1033) DLSERFRRLMRRADELARRGNPEEARKVLEEAEELLEQYGSPELLESVRMLLEVLG ( SEQ ID NO: 1034 )

[0407]

[0408] DISERMRRLMRRADEWARRGNPEEARKVLEEAEELAEQYGSPELLESVRMLLEVLG ( SEQ ID NO: 1035) Table 9. TrKA binders

[0409] SEQ Sequence

[0410] ID NO:

[0411] 16 RDEIKERIKKAVVRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIRQVEKRLR 17 RDELKERIFKTIVRAWTGDPELLKEAKKLLEKLKKLGRLDQSAKQLEKAVRFVEKQLRS

[0412] 18 RDEIKERIFKAWRAIVTGNPEQLKEAKKLLEKLKKLGRLDQDAKKFEKAIRQVEKRLRS

[0413] 19 DEIEKRLRTALEELVRADKNNDEEKVRKAWQKAVQIVIEANDDNVSKLASKIYRELAKRVKS 20 SPEEELKKLEKIAKKKGDTRALSVIQVAKKAYKKGDEWTLRIALHIAKQLL

[0414] 21 DEIEKRLRTALEELVRAVKNNDEEKVRKAWEKWQIVIEANDDNVAKLASKLYKEIEKRVKS 22 DEI EKRLRTALEELVRAVKNNDEEKI RKAWQKWQI 11 EANDDNVAKLAS KLWREI EKRVKS 23 DEIEKRLRTALEELVRASKNNDEEKARKAFLKWQIVIEANDPNVSRLASKLYKEIEKRVKS 24 DWKDQASELLDKLIKKARSNRDDELAKEVLKLLLRARWASHLSDDERAKKLLKKIVEEARKM 25 SDEKLKRLEKIAKKKGAWKALVAIHLAKLASKRGDEKTLRSAEKLAEKILS

[0415] 26 DEEEKRI LKKLKEVLRS LS S I PDPTAQKWETAWKLAVEAVI KNDKELLKKVAEYAERI LKKA 27 DESLKLLRKADKGAREKQISEKELKKAVEEARRRGLEEVAHYLELLLWVLRLL

[0416] 28 RDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDAKKFEKAIRQVEKRLRS

[0417] 29 RDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRLDQDYKKFEKAIRQVEKRLRS

[0418]

[0419] 30 RDEIKERIKKAWRARVTGNPEQLKEAKKLLEKLKKNGRDDQDYKKFEKAIRQVEKRLRS

[0420] Table 10. PDGFR binding polypeptides

[0421] SEQ Sequence

[0422] ID NO

[0423] 1036 DVQQEAKFVIARLAMLALHVDDPEQRKTVFKLMLLLADAVERGDRELLKKVIKTARDVISKVNS 1037 SEIEEVLELIRRYDPEWKELEKVLRALKRSGVSPEKILKYLLIVAHFLGSPLAVRLLFRLMTKR 1038 SLEELERLIRKLVKEGDPAARRALLVLESLKRRGVSPEEIVRRLIAFLFILGNPELIRLAIRLFL 1039 DLLEKAKKVAREILDPEWRIFELAVEVLKKKGANPSSILIAVLRLLRRRGVDPELLREVFRRAR 1040 DEREELIELILRNLDDPEKVHELLKRLLELAEELGDRRLYRYVMLIFWALRRNPERAERLLRKLK 1041 PELIEEALELLKEGDPKKVFRVLARLVQLLREKGDPRYRLVLLILAYVRFGNPEAARQLLRIVLK 1042 DELREILREALEKGDPELVKELLELLARLADETGDPKLRIVIAYVWFAKKKNDPRLLKTALQVLK 1043 DDERLARLAFRVLIKRAGVPDFDVKVTNGKVRVTITGRDQASQEALQLVFALARRLGLQVQIDTR 1044 DDERLATLAFRALIKRAGVKNLDVKVTNGKVRVTITGRDQASFKALQLVFALARRLGLQVQIDTR 1045 SELAEEILELIEKGDPRVRKLILELLKELKERGDEKRFKLLVRVWFLLRSGNPELAKQLLKKIKK 1046 DKAREFLRAAAKYGDPSAKRALKLLEELRKRGIPPEEALKIWEFLRKAGNPRLARWEEALKDE 1047 SLIEEILKEVRKKNPRWYILERLLRRLKEDGVPPEKILRILIAAAKRLGDPEASELLRRALKRK

[0424]

[0425] 1048 SVEELVKRILELLKEDEHRALRRLWLLLNRGISPEEIARIALRVLKLFKRDPEKARELIEKVLR1049 SELFREAIELLEEGNPEEARRVLLRLAIRLAQEGDPRRRAVALAYFYLKTGNPKEAAQLLEEARR 1050 SLKRIWILLLEAGVSPRDIARLFRIIDELKERGVSPEEILKILIKILEELGDPRAAKLLRKLLK

[0426] 1051 DETEEKAKRVSQYARNLGIEDVSISFHDNRLRIRVGGDDEKARRARRQIVRIAIQLGFRVLVITK

[0427] 1052 DLRDEFFRLLLTAIEIAKNVPDPRKASKIRILVAHVYFKARTLSKEQRVKLARELVDKAKKE 1053 DLSETVSKLASEVAKKVGVPSWEVRVHNGKVIVTLKGQDRAARRAMLLIRRIAEKLGVPVRVLVL

[0428] 1054 DIVELLTTHAERVATKLERSPDPRNKKVAKFIKTLIKKAKSVPDKDKSLRILAMAFFFAKRF 1055 NEELEKRVEELIKRIPNPKLRKVLRLLLKRLKEGDPKAAALLEILLAVFERGDPFRLILILKRVK

[0429] 1056 DKQHRAVALIRLVARRLKVPDVSVDVRNGRVTVRFGGTDERVRQAFLLARLIAKHVGVPIDVKVK

[0430] 1057 GPLERTARELAKELGIPPEDIERVLEVLERILGDPRRALWFLASLLTRAKKDPYVRELLKALIRL

[0431] 1058 SEQETASRLIKEAADRLGVSDVKVDVRNGTLRVTVRGNNESAQQLATFVLQIATQLGVRVLVLFR

[0432] 1059 DKERIFARALRALARRAGVPSVRVEVSNGKVTVSVKGKDRIARLFAQLVYNLARRFGIDVDVQLR

[0433] 1060 DDWRRILARALLLAKELGVPRWWRIDGRWWLVKGDNESAKKLITILKKWFEKKGDEVRVQIS

[0434] 1061

[0435] DVTEKLKKLITEAAKRAGVPSVEVTELRGKVLVRFRGRDRSVLRAALLARRLADKFNVPLRVI FH

[0436] 1062 DKLATRAIRLALQAHLQARQRGTPVRIHLSFKGFTIDVTVTEDPRSLERAIRLLILFLRRSSEN 1063 DPTSFRLVMEISLVLQLFKKGEIDPSQAKTLLKRLQEKAKKIGDRFAWLAEFAIRVIKKAEKS 1064 SLERIVRAAIEKAARRLGVPDVKVSVRNGTVHVKVRGSDRAALAAKIFAWNIARRLGVPVSVTIR

[0437] 1065 SREKAFASYVRQVAKRAGVPSVDVKVSNGKVTVTLDGDDERARQAELLIRWFSRHLGVPVTVKKK

[0438] 1066 SDDQEKLERIVHAIVRHFGLSPEWEKRLKKWRELLKKGVDPEQIVKIWKIAIRVHNQK 1067 SEELVKKVEELVKKTGDPLLKLLLEILKRLLKLNTPLARVLIDLILLLIDAGDREKLERAIRRLR

[0439] 1068 DKELLEEAIKLLKEGDPEKARKVLLELAREAHERNDPKLYRAIALVYFYLRSNPRLAIQLLELLT

[0440] 1069 DVERLVRAAVRKAAKKLGLPHWHVSVSNNTVRVQVRGRDKAALALLIFAWNIARAFGLKIELRHQ

[0441] 1070 PRRIEDALRSLLNSPNPELRLLAKVAIELLKRGVSPRRIALIIRRLADRLGLSPRVLRLVLIAVL

[0442] 1071 SKDEKARRLVLRILLEVGLKRFWLVRNGRILWIRGSDERAQTATKLVRTLAKKLGLEVRVLIV

[0443] 1072

[0444] DRRRRFI SRVLTIAKQLGVPRVI I IVNDGRVTVRVRGDTEKARTVQKFAQKVAEELGLDVDVQVR

[0445] 1073 DKLAVRAIRLALQAFLRARLRGTPVRIHLSFKGFTIDVTVTEDPRSLQRAVRLLLLFLRRSKEN 1074 S FEEVI HRALMVAFVEGDKKAFRI LTKI LELARQGKMS EEQAEKRARRI EEKLRK

[0446] 1075 DERKKVDAFLRFVIVRVFFVKDPETKKDILRKLQEFAEQISDEEAKKKAQKVWELLTRLS 1076 SVEERIEELKKEGDPKKALILELALELLKRIPPARVYALLLALARRLGVPPRDVARLLFIALQAK

[0447] 1077 DRALEIARSVLRIANEAGVPRLRVFVSKNTVRVRVEGDDDAAKRAKKLVEEYAKKLGLKVTVTKK

[0448] 1078 DYRTLFLQIWMVATTLAKEAKSEEDVKKAEKKLQEWLDKLNVNKLLQAIARRILRRVYKQLS 1079 DKDLKELYKELRKKLSPEEWKIVFKVIEEKFGDPRRAFAALYLVLRSLGIDPEEVRRLIRWAFI

[0449] 1080 SEEELIEKVERWGDPALRQMLRQMIEWLKKQGVDPKEAWLAAFALARHLGDPELRKVAREVLK

[0450] 1081 DPTRHRLLLRIFMVLVLFKKGKIDPSQAKTLLKRLQEKAKKIGDEDAVFAAKQAIDFIKRAEKS 1082 SLKELAREARRRGNPDVAFLVMLARYARKLGLPPKTIKKLLERQAKRRGIPPESLEEAKEIVEKG

[0451] 1083

[0452]

[0453] SERVAKIALLAALLGDPRYERSVKERVKELVKRDPRVEELVRELIKRLREAGNPRAARILEEALR1084 SLEEEIERALKRAGVPPQLIRIIKKLIRELEKRGVPPERIKRLVFTFLLQYDRRLALRVFFLLRK

[0454] 1085 SEDETFKKLVKKIAKEANVPSVRVTKVRGNFLVEIKGRDKAALRAALLIRRVAERLGLKVRIIMR

[0455] 1086 DELREILEKALKTGDPRLLKLVLELLMRLALKGDPKARRLLAFLQVLRRRHDPEEWKILLRLLK

[0456] 1087 SVLEELLRLARELGDKRLADIIRALIRAVRNGKISPEKALRLILAYAHFLGNKELFRWLRLFKK

[0457] 1088 DKLTKAAKELAKKVKSGKLSPKQAMRLLFVIAFRLGIDRKEMVRAAQHLMLLITK

[0458] 1089 SEKEYLERVLRLLEQGNRRRAIARLLQILQELKRRGDPRVEEFRKLVRLLARDEEKAKELVRRLR

[0459] 1090 TEEEKLKTKIKELAKKVGVPRVKVRVRDGRVLVLVIGDDENAQTFSKLLLRIALREGIPLFVFIQ

[0460] 1091 DNKVFMRAYFIVQDIQKKYPDPREAKKVAKEVLKRIVDSIDDPDERFWHAAARLLLRKIG 1092 SLEEALEIIEKELGNPKLVQALRQLIRLLKRRGFSPELILAALRLFLRRINTPEARLAYKILRET

[0461] 1093 SEESKRAKELIEAAARRLGLDPSAVEKARKLFDKLVKRGISPDEAFHIAIRFLMKLSTSK 1094 DARLRLIFWARFAARSAGARRVRVTEHNGTVRVEVDGNNERVQQLLKSVKRLAEKLGVPLEVTTR

[0462] 1095 IEEAKLLKRMVKAAAKRLSQGRLDEEKWRLIVIMASSRLFSLGISREEIRDFQRQAKELKKKA 1096 DERTKKKAEELFKRLIKILQNAPNPFLRQLAKELETKFRRAKKNNDDDTLIKLVFFAIMWRAG 1097 DERVRRIFKVLRELARISRNLSEEESERFFKKILEEAARRNDRILYLAAKLALRDSRRALHF 1098 SELDKIVKKLTDIAREAYKHNDKETIEKALRWAALFARQYRDPKFRELFRIIFQLRQRVER 1099 DERKKVARAVWEIAKRAGVKGFFVFVKEGKVWVTLRGTDESAQTAKKLIEEWSRKNNLDVTVRVR

[0463] 1100

[0464] DEKRRIFMRILLAARRVGVRRVLVFERNGRFKVFIQGNDERAKTASKLVTKLARKLGLDVKVTW

[0465] 1101 SVDELVEEAIRRGLPPEELEKLLEELLKKTGDPRVRYVIVALRILLRLGAPPQIVLWWLRIARR

[0466] 1102 DERREELVKRLKKSPDPSEKAIAAAYEYFLKMGLDPEMVLYQLKRFARKVGDPKLEELVKRLEET

[0467] 1103

[0468] SRLLRFALLVRRLAEDANVPHVRVWSNGRVRVLVRGNDESAQKFIKLFKTVATKLGIDVRVHW

[0469] 1104 THREILKLILKIAKKDEELAKRIYLIVWALRRGAIDPTQVERLLEKLIREFGDPLVRLVLELVRK

[0470] 1105 SLELVEEFLKKLNTPEARILLRALEFLKRRGWPPRLWQILYTLARKLGNPELAEEIRRLAREAP

[0471] 1106 GPEVLKKFEELRKRDPREALLFVLRIVFELGDPRLKRLAVRAWFLLKDGNPLVEQVIRRLRELLH

[0472] 1107 SEEEKITKTIETLARRAGLDDVKVNVKNGKVIVQIRGHDRAARRLAFLVIRWADEQGLKVLVQIT

[0473] 1108 SEDEKIRKTVEQIAREAGVESVRVNRINGKLHVTLKGDNRAARRVAALVIRYADEVGIRVDILLL

[0474] 1109 SSRRRVISRALQLAQKAGVKRITITEKNDRLRVTLNGNSPSIQKARRLIEKLAKEAGVEVEVRTR

[0475] 1110 SEEEKVAKVIETIAKEAGVPSVRWRVRGKIWIIRGTDEKARRALRLIFQFANKQNLPIYIRLR

[0476] 1111 DRSEVRKVFTLAFRVARLLASEDPNNRKRAKKEASTIKKELSKKSPSLSSLFEFLVRRITSRLG 1112 DPVEEKFKRLEKELRKSIKKGKISPEQALRILIKFMMHLFVRGKLSRNVAEMAVKFMREVDERKR

[0477] 1113 DVEKLVKEVQKLAKKRGDNRAHVLAFVILLMYRRGLISSEIASRVLESVAKRLKG

[0478] 1114 SEERKTLARLLLIAKRAGVPRFIVLHVNGRLQIIFKGDSPSVQRARKLAEEFSKKSGIPLKVRVK

[0479] 1115

[0480] DLDETVKKIAEDVAKEAGVPSFRVRVSNGKLTVRFKGNNESARRAFMRIRLLLRRLGVPVIVQW

[0481] 1116 SRVKRLSVRIVMLLLFARKAGLIDPEEVRRLYEEVNRLARKGDWKQLQKLFRKILEEIRRR 1117 SLLEELLELLKRYVSPELVERVERLLRALLRHGFSPQLVILSVIRLLEARGDPELARLLREILRR

[0482] 1118

[0483]

[0484] DQDRRILEIIHQIAEEANVPSFTSYVSDGIVFVRVWGNDEAAKTALTRIKKTAEKLGVQVRVRTL1119 PELRRLTARLLFLWQAIRRNDEDTVRTLLKIAKELADRNPKYKFVFLVLRSMIFRMRRRI 1120 DKSETFSKLAKEIAKKVGVPSVKVYNIRGRLLVTFRGQDRAARRAALLVRRLADKLGVEVRVLIQ

[0485] 1121 SRHRLVSQLIRKAARRAGVPHVDVRVKDGTVSVTIDGTDVAARAVFAFVKYVSKLLNVRVSVTFK

[0486] 1122 DEELIERALELIERGDPSVRKWAELALLLARKGDRRRLSYVARVLRLLKRDPRAAKQALKILLS

[0487] 1123

[0488] S RLLAL S I FI WN I ARAAGVP S VT VRVKNGK I TVTVS GNT E S AKLARRAVEAFARRYNVDVD FQ I R

[0489] 1124 SELLRFRLRLRRVADRANVRRVTVFRLNGKIFVLIKGNDDSAKIVEKFVRTVAKKRGIPVWKLF

[0490] 1125 DEKEKTVRSVIKRLQELSKKASDLRKRMRLIMLSKILTEALKEDKDEAHKLAQRVKEMLDKD 1126 SELKEWERVLKGELSPSEVLRVLAELYRKTGDPRYRNLLARLYLARRRGDPELLKRILELIRRA

[0491] 1127 SRERIARLMMIAFEKGDRRALVWAQAWFALRRGDEKTAKRLIKLASKLLS

[0492] 1128 DKETAFAALLRAMAKRAGVPSVRVEVSNGKVTVSVKGTDDAAELFRLLVMFLAKRLGIDVDVQIR

[0493] 1129 DDRDKLARLVIKAMELADQIKDERWSLHVKTLIAHAWFRNDTKLAEEVLRLAKKLVESVQ 1130 SERKTLYQLVARLAQELGVPRVLVRLSEGEFLVFVFGRDESAQTLSKILTTWAKKVGLKVEVILR

[0494] 1131 DEEVSRAYRLAAKLGDPRLAALIRLLLKLVKRGDPNLRLWAVARRLAKELGDPEILEVLEKLLK

[0495] 1132 SSEEALKLIKELAPPEVRIAIEALHRHLKRQGLPPRLIVQIIIKFLERRGVLPPEVAKEARELLR

[0496] 1133 DRDEEFQKKVESAYRRLKERNASDKRILSYIARLLREAQKIGATKARKLAAEIALRLARRLD 1134 HEEIAKKLKKKVRTILDQLNASDEQKVRAFFLLHFAERVLSRISDKRHAKRFADFVTQMISNM 1135 SEELEEVLELLKRDPKKAFRLLAELAREAQRRGDPELFKVLMLLLIYIRLGNLKLARQLLEVLLK

[0497] 1136 DKRQVLRVFEKAFHHMRHHPQVKKLAIKVMMLAVQAKKQNNPELLKQAEELLKEVEKRAK 1137 SAEEVLELIKELLPPDVRRFVERFIRKLREDGVPPEKIVELIRAALKYYGRLTPEIERLVRELRK

[0498] 1138 DWKLVIEVQKLAKKRGDLRAHILASRIIFAFMKGLMSSEEASKLLKTLAKRLKG

[0499] 1139 SELEELLERIKRTGDPKLIARVARLLFELAEEKNDPKLRRLAALVYFYLRTGNPKAAKQILKRIE

[0500] 1140 DERRRFIARLLQVAQEANVRRVLVMYHNGSIRVFINGNDDSAKIVEKFVRTVAKKRGIPVSVTTR

[0501] 1141 TEARRLARLIAEYARKAGVPHVWWLNNHVFVRFDGNTDSVKKASEFAKEVARQLGIRLRWTR

[0502] 1142 SPETLKLALFIALVRRLFKNGEIDPSQAKTLLKRLQEKAKKIGDHFAVRLAEHAIRAIKKAEKS 1143 SDELSKKLEKKVKDALKKGDKRLAKFLVIKAILEAKRLNDRKLEHVAARLALLIFKWS 1144 DLDEEVKKTLKKLYEIAKKDPSKKDEVEKLLVKVAAHAHFRIQNPEKRRKALELALTLMRRIG 1145 S FEEVI RRAI KLAEKNGDKHALVLLKVI LFLARQGKMS EELAERVART I EKRLKS

[0503] 1146 DKEEAERILSKIHELARRKNDRKVQRLVMRLYFALQFADKKNNKKMVDDITRLAKELLKKVES 1147 TKEKLREIVRLLREGDPRARELLRRLLKELAERGDRRLFFLVFAILRLLKRNPEEATKLLAALAR

[0504] 1148 SDESRKLQRRLALIFAALRQGFISNDEAKELAKRVEEKARKLGDKHVRQLVKFVKDAIERAG 1149 SLEELEEILRRLARKTPAAAILLRILERLRKRGIPPRLVLRALLLFAKHLGDPELEKLIERVLRK

[0505] 1150 TRELLLRVLILLWQLRTPIARAIIAFIYIAKKEGVSPEEILKIILKLARRAGDEEWRIIEELSP

[0506] 1151 SLLEELEELLKKLNTPAARMLIALLRRLLEEGRNSPKLILLFLWLLARRLRDPEWKVIREILRK

[0507] 1152 PVYVFI SNVTPEAEKI VKKIARLGRVLVI KRPGNRI FVYVQNPELAKQLVELVRKAGARVLI FR 1153 SELEELIERALKTKDPELLKKALLLAARLLDETGDPKLRIWAYLWFAKARNNPKLIKRALKAAR

[0508] 1154 SERRTVRRLLMEAALRLGVPFVFIRTVDGRVYVFVRGKDESAQTLSKLAEELAKKLGVKLHVRRR

[0509] 1155 PLEKLARHWTKAKEIVKKAQDPRLSKIFKEIADKLETALKNNDEKTVRYLIAKFFFLKRRLS

[0510]

[0511] 1156 SSEDMRKVFIMVLILAHEAAKDEDLERKARKKLDKILKDLKVPNAHFFKQMFDLLVKKVRSKLSDREEKITTLVKEAAKRAGVPHVDVNFSNGRIHVRVRGNDRASRRLFAFARNLALAYGVQLQVLNR 1158 DPTRHKLMLNIFLVLVLFKKGKIDPSQAKTLLKRLQEKAKKIGDHDAVHVAELAIVRIKQAEKS 1159 NRDRIRAELLLWAIARRLERNNLDPRLVRTAIQWARRLQREIQNPENRKFVQTVAEELIKRVM 1160 DRRERQLKLLIDFLVTRIKKYNPELAKKVESIVKKAEDNQDPNLLVKALLVAIKALKTVSS 1161 DPKKLDELRKKIETLLRKVTDPTAYRLVMRALFLAIMALKQQNPLLEQLARELYEKAKKIQ 1162 PTEEVIRTVMLLVFMARRSQDPTRVEKLLTKAEKLAKNINDPRLEELVKIAKKQARSILS 1163 DLKVLSRALLIVLKIQKKYPDPREAKKVAKEVLKRIVDSIDDPKERVRVFFFAHMLLRMIG 1164 SELAELLEEALRRNDPELLRRIVRLLARLAEKREDPRLWRLAALLWFALRTGNPLLIKAVIKKLK

[0512] 1165 PEELRKLLELIERGDKKKVFELVRKIAEKLKKKNPSLVRLLVAYVFLALRGSPAARRLLRSLAEK

[0513] 1166 SEEIFRKLMFAMFIAKRTGDPRFKKIAEKLNRILETIAKRDQDPDAKRLLRIFRQAAKRRKKS 1167 DLNARIFLLILRAARKNDEKLARELLDKSKKDPQVESAVERLRWFDPRVDKAYKILEKWS 1168 DKKLREELLRVALEAMKKIKDPDTQKSILKIAAQAFFRARNIDDPTERDKVLKKWLKLLKDFIS 1169 DEKAEKLILKVKMFILMIKLAQSKEMKSKASEAAKRLLTEISKIPDEKAQKIVKKLKEILDKLS 1170 SQLEELIERALKTKDPKLLRHAIIVAFLLLRRTGDPKLREVLHLLAEAFFKKDPELIERALRVAR

[0514] 1171 RYTEHVFFVWEATDEEARKRIRKRVEQIEREAKEEAKKRGREPQTYFVERTRDGYVSAVLIIIVE

[0515] 1172 DDRRTELLSRLVKKLLEQARQISDPRQARKISLKLHMVKNLAQNPKVRTLADRAIKQVNSIVRS 1173 SKLEEVERLLRELAKKDPYAYILLRLLKKLKELGVPPERILRLLVAAAKRVGNPELAKLIEEILR

[0516] 1174 SVLFEVAMVALELGDPRLEILVIRAILLLKRGDPRVKEVLKKLVELLRKLGNPELAKKVEELLKK

[0517] 1175 SKHEKARKALAMLLQIAVELGVDRRYMVRAMFIFLELVKNGVDLDQARRIAEKWLKKWKS 1176 NRADQLIYFVRKIAKRLGAPDVKVDVRNGRVTVTIKGTDFRARAVAAAARIVARLLNLELDVQIR

[0518] 1177 SYVERWELLKRLGDPVARELAELIEEILKRNYMLAMFAAQMAEEALKLNDPDMLDQLIRFLKHH

[0519] 1178 SAAARVALLVRRVAEKLNVPNLRVFVLNGKVQVLIRGDDESARTAYKIVKKISDTLGAPVRVQIH

[0520] 1179 DKFAIVMHLLFLAKRASKRNDKLLVLFVLRLAKRLYDKHPDDEKIEKVLKIVEKIYRKVF 1180 DIEEVLERLKELIRRGDPRVRRLLARVAMRLRQRGDPRARRFFLAFFRFAAGNPEHALSVIRKLP

[0521] 1181 SELEEEFEEALERGTVEEFLRRLYRELAREGDPRSAALVLAAFRLVKEGDPEAAKLALRIIERRR

[0522] 1182 DPTRRHLMSAIFMVLFLFKKGKIDPSQAKTLLKRLQEKAKKIGDREAVFLAELAIRIIKKAEKS 1183 DDRLRRELFLVWRVARRLGLDRHAILVAALLAARLADKGVDPTQAERILEKVLRTIKKRKS 1184 DEEDKVRTTVTEIAKKAGVKGFHVTVRDGKVRWLKGTDESARRAFRLIFQARHKNNLKVFVLRF

[0523] 1185 MEKYHRLLTKLSRLVIRAQQIPDKDKARSLVKTVAEEAIKSIDDPTLRIRFFFFVRFILENM 1186 TLKEVAQLLQEILQHVPDPRVRRLILIVYLALRKGNPELAERALEEVIKRLGDPRLREALKLIRR

[0524] 1187 DKDEVLELAQRI LKEVQERGNEEYS RRI LALVAAAFTAS RKNDEDFLKFI LKVI RELYKKLQS 1188 NLSKHVAAAARIIARFVGVPSWSIDVKNGTVTVSLKGRDSRAQQAAYMIERFAKRLGVPVDVQVR

[0525] 1189 SNSEELAKLIRQVAREANVPSFSVDVRNGTVHVTLKGDDESFRRVFVIALQLAADLRVKVRVTVR

[0526] 1190 GPEEVIEIIAKALGVPPEKVRRAVELMRKRLRIPPERALWLLLNLVKRLDPRFKEEVEEARRLLQ

[0527] 1191 GEEEALKILAEVLGQSPEKIRAAVRHIRRRFNVSPEEAILLLAKLIARHLNDPRVAEEIRKALEK

[0528] 1192

[0529] S DRKI AARAI RI VARRAGVP S VEVTTTNNT I RVTLQGRDEAVRLAAI FI WNI S RRLNVPVEI RI R

[0530] 1193 DELQKIATRLIKLAEKKGDKRLAMIARAVKFMIKRGELSKEEATRLLRQIEERAS

[0531] 1194

[0532]

[0533] SEFREVIRLLEELGNPRVARFVRLAWWLLRLGNPENAKAALKAALRLAERLGDEEAKRLIEKLLK1195 SARKKITRILLELADRGDPLRKHWRVWFILRKDLDKAEKLVRQLIERFGDPLLREILELIKELK

[0534] 1196 SSREELKKLVEKIAKKVGVPSVTWERNGKILIVLKGHDEAARRAALLIRRVAEKLGVPVRWIR

[0535] 1197 DWRHAALLLAVIARIAKQDNDEDLLKRARTLFELLHRAAQRGDEDTVKRVIEEAKKLAKDL 1198 PLESMARFWKRAKEIVKKAQDPRLSKIFKEIADKLETALKNNDERKVRILIAFFFVLRQHLS 1199 DDRTARLLSKWKSLLEQARQIEDPQQARRIQVKLFMVKRFAQDPRVKTLADRAIKQVKSIIKS 1200 DQEELEDMIKAVEKFAEHLERNPDPKKKKAAKIVKDAVKNAKQIPNPRKAKKYLMIWFRLRKS 1201 NVEELAKKVQKLAKKKGDRRAFLIAFVLVRALRKGILSEEFAKRVLEFMEKRLLK

[0536] 1202 SEETVRKAFQVATRRGDQKAARTLMRILFMLRMGVLDEEEQKKADKYAKSVIS

[0537] 1203 DEFRRKAARLALRHLEILRNLSEEEAKKWARKILQEAAKKQDSALWIAARAALDFWRRALKD 1204 SERLELIVRTLAHLIRFADERLRKLIFKFLLQLHEDELRKSSHFLKRASDEEIKKTAEEIIRRA 1205 PIEELIKELSKRVPPRLRQYLEAALRLKKRNPETARRLLAIVIYEILRLDPNDEIARKVLRLLMR

[0538] 1206 SETSKKVREFLELVIQRGDERKFMMMIRFAKKRGLLSDEELDRWRKRFEKVKK

[0539] 1207 SIEEWEKVLKEAREKGDQRWLLAEALKRALERGDKRRALALLIILRRLVGDPELRKAATLLVR

[0540] 1208 LERDILTAFIIADELKQEFKDDPHIKSLVHRALIELMRAWRENDEDKIKEVIKKIKTIKKKADS 1209 SLLERIEEALRRNDPRLVLLLLAELLHLTGDPRVRFVLVYFYFALKEGDPEKAKELVKELLKRVK

[0541] 1210 DQMERLRKLAAHIVMLAQKVGLSPEIIHRIARKAFQVFQDRDEDEARKWVEEVINRWKKKL 1211 TKEALRLILRLLDKRPDDSLLRRIMAYIFIAKRLGNPEFVERIVRQLLKRTGDPRLKEVLELLKK

[0542] 1212 SEELVERAVKFAGLSPHVEKILRAWRVLERLGASPQQIKYILTRFLRRLIPDPEKLRRAEEILR

[0543] 1213 DEELKKLLKIWRLLIEADENNDEKSARLARRAYLLLWLAKRADEEHQRQLKTLVKQLIKKIK 1214 SLELVEEFLKKARTPRAFLLLRLLERAKKQGLPPEKWELLRAAAKALGNPELAEEIRRLFEEAP

[0544] 1215 DIEEALELARRLGAPPRWDYVEKLARRLKEKGLPPRWLALALYIARLLGDPLLEEAIKRLLKK

[0545] 1216 TEIKKIAAAVFRALREGTLEEFLRELLELYRKKGDPRILHRLLIAISIARTLSPEAREIVKRVRK

[0546] 1217 DDRVKKARKIASTILRRLGVDEAEIRAAEAIVERLVKKGVDPKDAARVAVLAVRFLLRIRR 1218 DAIRRIAELAHRYGDPLLKLWAAVWLLKRDPNDRAARHLLRALEQAAKKKGDPVLEEAIRLAKK

[0547] 1219 PEIMVMMVEKLITLLKKLAQNPEMRKDIERLEKEIRTRIKKDPRFARKAFMILLQLYHFLKQQM 1220 PHREVLRLLAELLERARDPRQRSLLRLILFALHLRNEEMLKQLLEALERVLGDPELVRKLEELIK

[0548] 1221 PEARQITLRLSLEVARAIDRNDEDTVRTLLKIAKELARRNPQYREVFILLYNFIHRAKRAL 1222 DVEELLEELERLGLLDPELLELLKELIRRLKEEGDPEKLRKLHLLIAMARKDPLARFLLHFLVRK

[0549] 1223 DEARSLALEVAIRAILKARQLNRPYDVTFTVGGQKYTIRVDTDPSKQRRALRRILLIFMRYAFK 1224 DKTEIIREMLTELAKKIKSVSDPTQKEKLKRHLFLLVFILKNAPDPTEREAAKYAETLIKRILS 1225 DPRHEALRRVFSRLYLLLYRLTNPEEKKKAQKWAEKVLQEAGVDPQIARAAVEIVT READ RAR 1226 DEELKTLRSIVETLLEKARRNNDERSRRLARLALFLLLRARRADEEERKVLKRAVKALIKAIK 1227 SDELSKKLEKKVKDALKKGDKKRAHLLWHALLEAKRLNDRKLFKVAAKLFMLIQKWS 1228 SEAEEISRLLRLAAKALGVPSVEFDVDNGTVTVTVHGDDDAAQILWSLVRRVSREEGVPVRFEFK

[0550] 1229 DETSKAVRMLAKVILLLKRNPNNEEAKKLYKEAVTIAQKVIKKLKDPKKQRHLHFLIHLATLFS 1230 DEVERVLEILKRLGVDPRLLQAARQLFRLLRKKGLDPEQVRLLVAVFLRRRDEDASRLFRKLLRK

[0551] 1231 SERLFFSLRLRGDNRTSRELAEAWARRLAERAREEARRRGLEIRVHFRFTRENGKLTVTLRLHIR

[0552] 1232 DVEKKAKEAI KLARKKGDKRAVLI LHAI AVMYRRGLI DKNDADKLI EFI I RRLRS

[0553] 1233 SAVEKAVKIIKKLAKDLNIPRVQVRYRDGRILVLIHATSPEAKTLRKLILEVARRLGVSVFVYLQ

[0554] 1234

[0555] SLEELERLIRKLVKEGDPAARYALRVLRLLKKQGIPPDFIVAILLKFLRRLGNPELIRLAEKLKN

[0556]

[0557] 1235 NYMRVIMRAVELALKKGDKKAVRILFHIAMAARIGTLDPTKAERLARRIEKKLRS1236 SLEEEVERLLREAGVPPKIIRLLKAIIRLLLKRGIPPVAILRILIKFLEKLGDPRAAKLLEELLK

[0558] 1237 SEDEKIKKRVKEITKLAGVPSVEVRIKDGTLTVTVKGDDRAVRAALARLLNLALLLGIPIRIHVR

[0559] 1238 DKEEEKILELLKQGNIEEAIKLARELARRLKDPRLAIAAAAIKHLLRRGAPPQIILYWRRLLEP

[0560] 1239 DEKEELEELVKKLGSERWFRMWYLIARRLGVPPEEVLKRIYELVKRLKVPPKSAALHLLQELNS

[0561] 1240 DAEKIIRAAVRAFARLARIPSVTVRKSNGKFHVKINGRDALAQQVAYAIKRIAKKLGLDVTVEYR

[0562] 1241 SESDKLSKRIKELLKKAKEKQDKRKAMLAYLLALRVLRRYPDDEKIREVLKEAREWRKIYK 1242 VPELERVKELVDRYAEKVGLSEEEKKKLRKLVDKLYKKGLSPMQVAIFVILFIRDLKKKL 1243

[0563] D VL E KVL K FVRKKN EQ LARAVEAL I RRL RE RGVS P RT I L I L LWT I L RKAGN P EAVE L VE E L L KRL

[0564] 1244 DELKKAAKELAKLVKKGLLSPLKAIALLIDIARKLGASKEEQRRALAALLYLIRH

[0565] 1245 SEVREILREAARIPDPREVKRLVLKAAKLAIKLGDPKLARLAALVYFYLRLGNPEAALHLLERIG

[0566] 1246 TKLLRALAILYEKTGDPKLRSLLLWFIELRKGRRSPEEILELIKKLLERLGNPELAEELERLFK

[0567] 1247 DELEELLRELRKDPRLRAAVLLFRALLRQGYPPELVSIAVRHFLRRNGVPPELIEKLLKILRDLG

[0568] 1248 SLQERFQRLARAAARYLGVKSVTVRVRNGTVHVTINAPDTRAYILLKIIKKVAEQEGVPVDVKIK

[0569] 1249 DVERVRRRIFWLWFAHQQKDPRYADLALKLAQTLVKLYPDNAHARAILREVKERVEKIKS 1250 DERRRLFVAIRNIALAVGVPSIRWIHRGKFRVRVSGDDDNARKALKLIKEVAEKLGLEVTVEYR

[0570] 1251 DVEKLVAEVALLALKRGDSRAMWKAIAILLNYRRGKLSSEEASKLLKTLAKRLKG

[0571] 1252 GPEELEEIVRRFLPPKWRYVRALIRALARLGAPPQIINYAIILFLRKQGLVSPEELEELEKLLK

[0572] 1253 VREEALRI I FKAI LLLMNGQI SRREAAKI I LLAREASKKNDEELI KKVKSLLDSVEKKNQS 1254 DLKQRVRHLMIRILLLARKKQDPRLRRLAARLALLAIEAQERNDEEKLDKVRKEAKKWDSG 1255 PPVLERLLELLKRLGNPLREAVRALLRALRRGEIPPEKVLKRLLFAAYRLGDKEFFKLVAELFRK

[0573] 1256 TRREAMFVLFVAHFAYRKGDKDMVHAAYEVAKKLLEKEPEDERVRKVLEIVEELLKKVS 1257 SAAEEALKLLREVLPPEEREILERILKELRKRFPPAAVIFLLAKILLKLGLIPPQVAAKVLEILR

[0574] 1258 DAQRTVEKVIRHAQEFVSKKGDERIKKQVDTLAKKAQTLLKQGDEDSALEVAALLMHLLWWKR 1259 DSEEAEKIIRRVLIFAI FQNDKKIFRSAFELLKILEEATKKGNEELIKKVTKIARKLLSYV 1260 SREDLFAWLLRRWKRLGVPSVDVKVSGGRLTVKVKGTNQDARLAAAAAKVLARFFGIEVDVKVK

[0575] 1261 DERRRFISRVLQIAQQLGVPRVLVLVKNGRFTVRVRGTDERAQKAKEAIEKLAKRLGLEVELKDG

[0576] 1262 GPLEELEEILKELAKTDPAAKILLAALKLLRRRFDPRLVLMILELVARRLGNPELLKIIREILRK

[0577] 1263 SHLERLIERALKTKDPRKLRIAIAYAWFLKERTGDPKLRRWDLLARALVEKDPELIERALRVAR

[0578] 1264 DEDTKAEKIAKDVLKDLVKKATTEREVLRALLHAIMVIRTLIPDQEKAKEVRRELAEIALKWL 1265 DREELKDLVSKVEELVRKVIKDPNYKSMATRLIKKAKELAKKGDEESAKRVIWMLIAIVFMYKR 1266 DVEELLEEAVKTNDPRLVREALRLLFKLLDRTGDPKLKRIIAYLLFALKKNDPKLLRQWELLKR

[0579] 1267 STESVKKALKKALKSGDARKAFRIVMLAKVFGLISEKEARKWRNLILKVSSI

[0580] 1268 SELEEELKRAIEEGDRRLLARLLLRLSRLLGDPRRAVRLLLRLIARVLNVPPEEAAKILRRLLKT

[0581] 1269 DPTRHHLLKRIFMVLMLFKIGKIDPSQAKTLLKRLQEKAKKIGDKAAVSLAELAIVLIKQAEKS 1270 SREELLEELEKKARNPELRDLIRLLKELLRRGDPKVRRVLFLLFFRARLNGNPELARLVMKVFKT

[0582] 1271 SEQHLREWIKKVEEQIRRGTITIRWAMIVHILIRQAQKLGDEEVEKRLRELLRRIAEVFH 1272

[0583]

[0584] SIEELLEKLLRELGVPPRQIKIVKAAFRRLRRLGLSPERALEVLTILLRQRGNPELAKAVRLVLK1273 SERHIKHALALLRRGDPKRALFVLYYLARRTGDPRVKELLRLLLEAFLKNDPELLKRVRRLLEET

[0585] 1274 NEQDKKFAKRLLDQAKKLADQISNDRLRKKAQSLVEKARKIDDPNVAMKVFVFVHLIYRHVQK 1275 SRKVIRELAMYVTRVAEKLGLDRRAFRRAFSLAYLLAERGVDPTQAKRILEKFLRTIKKRKS 1276 SLEEALEIIEKELGDPKLVKQLRAAIRVLKRFGADPFLILRILRRILKDADTPEARLALKILEET

[0586] 1277 SELEKILREVAKRVYKKKGDPRTIMKVIYQIVQRLIKTLGLDEEQQRELMKRAFMVWMDW 1278 SEHLKRILKAAVRFIARLLGLDEKAQKIAKKVAEKLADKGVDPKEALRLLIRVLQQLYVRLR 1279 DEVLERLRKLLRKLGNPHLVRWEAFIKYFRRLGLDPRMIIFFLLKFARRLGDPELTRVIEEAIK

[0587] 1280 SLLEELEELLKKLNTPVARLLIRVLRALLKRGRFSPKEILQILWSLARRQNDPEWKVIEEILKK

[0588] 1281 DELLEELEELLKKNDPKVRRLVLELLRLAEELGDPELFKRLRYIAAAVFLGNFKLALQVLKHILK

[0589] 1282 PFKEILKTLYKLIQEALKKNDRRKWRLYILLLRLMRRAQTEEARKLAFKLLLLVIDLG 1283 TEELEKLLEELLEKTGDPRVRILLRALRVLLRRLDPKQWQILWSVARRLGNPEAQRLIEEVARK

[0590] 1284 SLEELVKELASEALGRPPEAVEKALRHISERFQIPPKRAIRLLVRALLHADGDPRLKELYKLVTK

[0591] 1285 SELLRSARRLLAFIALALARGVHAEEIIRWLEEWLKRARENQNKQEIYSAQAALLHARKWLER 1286 SPEEKAKKLAEAAVKFLVKLASEQIAKYNVSRFVEMVAKDLNLDDSQTEKLQKYAQTLVHQLYS 1287 PYEEEIKKYADTLLTLIKNAKDPRQRRRLSKELFTTLLKLLKAPNPEKRAAAAQVIFKVFRVLK 1288 KEIAKKLADKLVTEATKLATSSDPNNRTKAHKLILRAMMFARFANDPNVYRKVQSVYQLIRKLG 1289 SVELIEEALKLLGDPDARILVALIRYLRKLGVPPRLVEQILEKAARQLGSEEIFRRAKELAERLP

[0592] 1290 SKHEVDLLVAFVMVIARTRGDPTARKIAAKLALEAMKAKDNNDDEKLKKVSKRLKSFIKTF 1291 S DEEDRLLKRALAYLWALKAGKI S VHLAKLAVRRVI EKARRLGDEKAVKVAKKI LEI LKKI EKG 1292 PEELYKRATKILTEAKKRAKDNPEILRAIIRLEFLLRVAYKQNNEKTLKAWKLVERKIRTLLS 1293 PEAKEVFRTLLEVLHKAREDGNKEWKLASALVAYFFFARKRNDEELLKKVITLAKKVIKKQ 1294 SPVKEIINRALQILQKLLRLGLISPEEAQRLALFLLLAAKEGDEELARRLYKKIEKIRRKLS 1295 DKEYVKRLVEELIKRGDPHRALMVLFVELQHTGDPDIRRLFRLLARVIIRYPDPREVLKFFKKLK

[0593] 1296 SKEEEAEKLLREVLPPRQRKLLEAILRILRKYAPPQLVIWLLAKILLRLGAIPPEVARRVEKILR

[0594] 1297 SEDEQLSREVEKTIKKAGASHVLVRVKDGTLRVLVRRPSPEADQVLNKLRQKAKKLGLKFRWFIF

[0595] 1298 SEELKELVERFERGEIPPKELFRLILRLLQRTGDPKLKRILAYLLFLIRRNVPPEEITRWLKAVT

[0596] 1299 SELLRVMFVAHFLARRAGVPHVHVRVSNGRIQVTLKGTDERARTVQKRLTQFVRAFGVDVDVQVR

[0597] 1300 SDELSKKLEKKVKDALKKGDKRRAKLLVIEALVQAKHLNDEKLHHVAVRLMFLIMKWS 1301 EEAREIIKLLFRAMQLASKKGNEKWRIIKHIMARLFVALRSNDEKMIEDASKKAKKVIKII 1302 PEEFLEEIKRLLEKLRTPAAWLILRVLRKLEELGTSPEEILRILRALARRLGDPEVLRLVEEALH

[0598] 1303 SKLEEVLELLKRVPDPRVHAWAAILALLERGRYSPRKLLLLLHFLLKRLGDKDALELLERLRRR

[0599] 1304 TEHEKVRLWEVAERLIRRGDQRRAQFVLWIAELLVRKLGDEKWLKIIEELKKELEKLKKK 1305 SVEEWELARRLGNPALAANLENIIRKLKRRGVPPRYILVALLYVARLAGDPEVARFIHELAKSP

[0600] 1306 SVAEELEELLKRGNAEEAEKFLRELVERHGDPRLRIALRLLERLKKSGVPPEKVIRFWAALRRL

[0601] 1307 SEVARRLLELLKQGNEEEARRLVLRAAIRAAREGDKKLAHLLALVYFYLKTGNPAVEDLLRELEK

[0602] 1308 PKERIKELARRIKEGDPEAKKELAKLFLEAVRRGDKRLFLAIIFILALVLRIPPREAARLLIRLG

[0603] 1309 DKDKLLEAAVRWARVAGVPDVKVRKRNGKVEVTFRGTNHQVQQASFMVKKIAERLGVTVTVRFE

[0604] 1310

[0605]

[0606] DERRRFISRVLQIAQQLGVPKVLVRVKNGRFTVRVKGTDERAQKAKELIEKAAKELGLEVELRDG1311

[0607] SLRLKVAFLLVKAGVDPRLITRLLQYIHQLERRGVSPEEILKILIKILEELGDPRAAKLLRELLK

[0608] 1312 DKEEVIKTVTKLLNEYTKRAEDPRKRLWAKAWKMLQDLFKKDPDEAEHLARRISETVKKKLS 1313

[0609] SELKEWERVLKGELSPEEVLRILLELAIRKGDPKYIYLWLWLRSALRRGNPELLRLILKQIRRA

[0610] 1314 DRRRRFRLTQQILTTALELYKRDSRQAEKFLKTLKEEARRHNDYALWIATRLAEREFKSDTHD 1315

[0611] SELMEAVFLARRLGNPTVASFLMQLLFALKKGRISPEEVRRILEELLKKYGDPALELALELLRRK

[0612] 1316 SRTDRWKRLREILKEAKEKNDPKLAIKAAMFAHVFLKEDPTDEKIRKVFKEALKVALDLLR 1317 SEIEKKLRKLFKKAHEKGDLRALAYVLFAIKFLRRGKLDPRAAEQLARIAERRLRS

[0613] 1318

[0614] S N S E E LAK 11 RQ VARQARVP S F S VD VRN GRVH VT LH GD DAAVRH VLARAYMLARRLN VKVQ VE I R 1319

[0615] SEDEKSKEISKTIQKLAGVDRVQVRIRKGRLLVFVRGSDERAQTALRLALEIAKRLGVPVTRHVQ

[0616] 1320 PMDTFMVELAKRLADEIKKLAQDPNLQKKADKLKKLVDEAYKKNDHITLRFWAMIILLWKRVL 1321 DQREILKVLREAWHTIRDASDRRVRAALRHLLVRLYEALRRKDEEALKKVLKRLKELIKEQK 1322

[0617] SVREAVEIILKRVGVSPEKINQLKANLWVLERRNPLAAEWLALKIIARVLNVSPKEAQRLLKELQ

[0618] 1323

[0619] DKEEKIKKLAERLARRAGVPRVSITFHRGRVTITVDGPNSAVRRLAAYIFLFAEKLGIRLSVRSK

[0620] 1324

[0621] NELFRLFEELVRRGISPEEILKLLVKLLREKGVDPQEIWRRLALILLQLGISPEEVARLVQKAVR

[0622] 1325 DKDDEFLDKVRKTYESLQRNNVDPNQIVKILARLSMTAMELGLTRAMIFVIQLFLKAKQQAS 1326 VREEAEKIVRKARTMLKKGLIDEKEARKIIFLALHAAKNDDEELIRLVKRLLEQFEKKRQT 1327

[0623] DEEFRVRMLIRFLARLAGVPDVKVSVRNGTVHVKVRGTDRAARTALQVARIVALVLGVPVSVTIR

[0624] 1328

[0625] SRDKAIRRLAELLGLSPEEVKKRLEELERKLGDPRRAVQVFLSELHRRGELTPRVRLFLLLIKRN

[0626] 1329

[0627] SKKEILEKVLELLKKDPKKALRLIIELLRRTGDPRVKRIAAYILFLLRKGDPAARDIAEKLLKET

[0628] 1330

[0629] DEVLERLLKLLRKLGNPQLVIIVEALIKALRRQGLPPRLIIQVLAAFARKRGDPELTRVIEEAIK

[0630] 1331 DFVIVFVFEGRRTRTFFFRSEEQAKELVKRLQEKNPRAKWTRHRGDRVIIFVDV

[0631] 1332

[0632] SKEDEISKTLEKIAKRAGVPDVKVTVRNDRVKVEIKGNDRRALAAYLLLLRVARKLNLKVSVLLK

[0633] 1333 DEKQQKLRRIAIRIFAFANFVGDPNVRTEVFRLAQHIMTWQRDPDKAEELLRKLMTKVRRLTR 1334 DDERLATDAFRSLIKRAGVKNLDVKVTNGKVRVTITGRDQASQKALQKVFALARRLGLQVQIDTR

[0634]

[0635] 1335 SEQDKQVKKLTDIQREAYKHNDKETIEKALRTAASFARQHRDPKFRELFRIIHQLRQRVER Table 11 EGFR binding polypeptides

[0636] SEQ ID Sequence of target binding polypeptide

[0637] NO

[0638] 1336 SDELFSKVFLKVTELSMIVMNAKTEDEKKTALTKIKQIADKVQDEELSKFVKRALEHVKKEVG 1337 SPDEAKKLLQEAEKLARKQNDRMELAYVEFLKHVLENAKRLNDKRAVESVRELARDALEELQS 1338 SLDEAKKLLQEAEKLARKLNDRMELAYVEFLKHILETAKKQNDKRTIESVRDMARDALEELQS 1339 SLEEVKELIRKLVPDPRLVWALEDLLELLKKGDPLAEEVLRFYLASAREQGDKDLTKAIELVLKT 1340 DDESLKLLLILVQIQLALERGEISNDQAKELAKRVEEKARKLGDEQVQRNVENVKEVIERFG 1341 DHWEEVFRWALELLQEATEQNDPTKAKKILEEAHKLLRRELSEEEARAWRYLKQLVDRELS 1342 DHWEEVFRWALEHLQEATQQNDPQKAKKILEEAHKWLRRELSEEEARAWRWLKQLVDRELS 1343 DEAAEEVLRYLKKLGDPELAELIERLLERVRKKKDPDLERTLEIIAVAVLYGDPEIARQALRALH 1344 DELRELINDAQNKNDPELLKKVLELIEELIRRGDPSYQEIVLLLLFVADLLGDPEITRLAEELLK 1345 SELSEEALRLLEEGNPRDAEHVLIDLLFELENRGDKELSELVLEILELLIRGDPDEARKLIKELT 1346 PLEEVKKWEEALKDDPELVRAVKTIIEAVKKGENDPIFLELLLRLLIETFGDPRLREALELVRK 1347 SDEERRELEERLERAVREGDEHHLRFLSQRLILLWWIAGKLDREEARKYVEELIKELKKRV 1348 DLEEEAEKLVKEAEKYARSVDDPAAKMHVHLVKLRLKLAKERISDKEKILRFVKEQVDLVKET 1349 DRHVADLIQHLLERGVPPEEILELIEDYIRWTGNPRFKKALELLEKLTRELGDPEEALRLFVERG

[0639]

[0640] 1350 SPEDAEKLIKTIKTIAKKRNDRMELAYVKFLESSLKQAQRYNDEERIEDITELLEDALKEIQS1351 DYTVDLAIKFVEEWKELKNDPDPRSQKLAEELERLIRKAKNAEDPNKKHTLVHKAFDIAVQAM 1352 DEEQETRLKLLLLLVQLQHLAKKDPDTAETEIDKIYKLAKKLGDETIRENAENAKRIVQELK 1353 SEEALKIFLAILRIQLHYERGEISPETAIRLLQKLIKRAEELGDEDVKRLAKRLLEKIRKEK 1354 DLDQLIETLIKLLKKLSKKYPEFKFHARYAIERANRIRKSDPRNSRAAVSLLEQALERIQKQA 1355 DYFFHSTRTYRGDGAEERAEKEAKEIVRKIAETIARKLNAKRVTLQVIWRRGDLILISLLFLIR 1356 DHWEEVFRNALEHLQEATQQNDPQKAKKILEEAHKKLRRELSEEEARSWRWLKQLVDREKS 1357 SLDEAKKLLQEAEKLARKLNDRTELAYVEFLKHSLETAKKQNDKRTIESVRDMARDALEELQS

[0641]

[0642] Table 12. IL-2RG binders

[0643] SEQ ID NO: Sequence

[0644] 1358 DVEEVFLRSARELIKAIERTGDPELEENLQQAMWALRGIKSEEAEELLERAEKLL

[0645]

[0646] 1359 SDVEEVFLRSARELIKAIERTGDPELEENLQQAMWALRGIKSEEAEELLERAEKLL Table 13. IL-2 receptor βγc heterodimer binders

[0647] SEQ ID NO:

[0648] 1360 GSHMPKKKIQLHAEHALYDALMILNIVKTNSPPAEEKLEDYAFNFELILEEIARLFESGDQKDEAEKAKRMKEWM KRIKTTASEDEQEEMANAIITILQSWIFS

[0649] 1361 HMPKKKIQLHAEHALYDALMILNIVKTNSPPAEEKLEDYAFNFELILEEIARLFESGDQKDEAEKAKRMKEWMKR

[0650] IKTTASEDEQEEMANAIITILQSWIFS

[0651]

[0652] able 13A. IL-2 receptor βγc heterodimer binders

[0653] G1 neo2 33 STKKWQLQAEHALLDWQMALNKSPEPNENLNRAITAAQSWISTGKIDLDKAEDIRRNSDQARR EAEKRGIDVRDLISNAQVILLEAR ( SEQ ID NO: 1362 )

[0654] G1 neo2 34 S T KKWQ LQAEHAL L DWQMALN KSPEPNEN LN RAI T AAQ SCI STGKCDL D KAE D I RRN S DQARR EAEKRGIDVRDLISNAQVILLEAR ( SEQ ID NO: 1363 )

[0655] G1 neo2 35 STKKWQLQAEHALLDWQMALNKSPEPNENLNRAITAAQSWISTGKIDCDKAEDIRRNSDQARR EAEKRGI DVRDLISNAQVILLEAC ( SEQ ID NO: 1364 )

[0656] G1 neo2 36 STKKLQLQAEHFLLDVQMILNESPEPNEELNRAITDAQSWISTGKIDLDRAEELARNLEKVRD EALKRGI DVRDLVSNAKVI ALELK ( SEQ ID NO: 1365 )

[0657] G1 neo2 37 STKKLQLQAEHFLLDVQMILNESPEPNEELNRCITDAQSWISTGKIDLDRAEECARNLEKVRD EALKRGI DVRDLVSNAKVI ALELK ( SEQ ID NO: 1366)

[0658] G1 neo2 38 STKKLQLQAEHFLLDVQMILNESPEPNEELNRAITDAQSCISTGKCDLDRAEELARNLEKVRD EALKRGI DVRDLVSNAKVI ALELK ( SEQ ID NO: 1367 )

[0659] G1 neo2 39 STKKLQLQAEHFLLDVQMILNESPEPNEELNRAITDAQSWISTGKIDLDRAEELCRNLEKVRD EALKRGI DVRDLVSNACVI ALELK ( SEQ ID NO: 1368 )

[0660] G1 neo2 40 STKKLQLQAEHALLDAQMMLNRSPEPNEKLNRIITTMQSWISTGKIDLDGAKELAKEVEELRQ EAEKRGI DVRDLASNLKVI LLELA ( SEQ ID NO: 1369 )

[0661] G1 neo2 41 STKKLQLQAEHALLDAQMMLNRSPEPNEKLNRIITTMQSCISTGKCDLDGAKELAKEVEELRQ EAEKRGI DVRDLASNLKVI LLELA ( SEQ ID NO: 1370 )

[0662] G1 neo2 42 STKKIQLQLEHALLDVQMALNRSPEPNESLNRMITWLQSWISTGKIDLDNAQEMAKEAEKIRK EMEKRGIDVRDLISNIIVILLELS ( SEQ ID NO: 1371 )

[0663]

[0664] G1 neo2 43 STKKIQLQLEHALLDVQMALNRSPEPNESLNRMITWLQSCI STGKCDLDNAQEMAKEAEKIRK EMEKRGIDVRDLI SNI IVILLELS ( SEQ ID NO: 1372 )

[0665] G1 neo2 44 STKKIQLQLEHALLDVQMALNRSPEPNESLNRMITWLQSWI STGKIDLDNAQEMCKEAEKIRK EMEKRGIDVRDLI SNICVILLELS ( SEQ ID NO: 1373 )

[0666]

[0667] Table 13B. JL-2 receptor PYc heterodimer binders

[0668] G1 neo2 40 STKKTQLLAEHALLDAFMMLNWPEPNEKLNRI ITTMQSWIYTGKIDADGAKELAKEVEELEQE 1A~ YEKRGI DVEDDASNLKVI LLELA ( SEQ ID NO: 1374 )

[0669] G1 neo2 40 STKKTQLLAEHALLDAHMMLNMLPEPNEKLNRI ITTMQSWIHTGKIDGDGAQELAKEVEELEQE 1B~ YEKRGI DVEDEASNLKVI LLELA ( SEQ ID NO: 1375 )

[0670] G1 neo2 40 STKKTQLLAEHALLDAFMMLNMVPEPNEKLNRI ITTMQSWI FTGKIDGDGAKELAKEVEELEQE 10“ FEKRGI DVEDEASNLKVI LLELA ( SEQ ID NO: 1376 )

[0671] G1 neo2 40 STKKTQLLAEHALLDALMMLNMVPEPNEKLNRI ITTMQSWI FTGKIDGDGAQELAKEVEELEQE 1D~ LEKRGI DVEDYASNLKVI LLELA ( SEQ ID NO: 1377 )

[0672] G1 neo2 40 STKKTQLLAEHALLDAHMMLNWPEPNEKLNRI ITTMQSWIYTGKIDRDGAQELAKEVEELEQE 1E~ LEKRGI DVDDDASNLKVI LLELA ( SEQ ID NO: 1378 )

[0673] G1 neo2 40 STKKTQLLAEHALLDALMMLNLLPEPNEKLNRI ITTMQSWI FTGKIDGDGAQELAKEVEELEQE IF- HEKRGI DVEDYASNLKVI LLELA ( SEQ ID NO: 1379 )

[0674] G1 neo2 40 STKKTQLLAEHALLDAYMMLNMVPEPNEKLNRI ITTMQSWILTGKIDSDGAQELAKEVEELEQE 1G~ LEKRGI DVDDDASNLKVI LLELA ( SEQ ID NO: 1380 )

[0675] G1 neo2 40 STKKTHLLAEHALLDAYMMLNVMPEPNEKLNRI ITTMQSWI FTGKIDGDGAKELAKEVEELEQE 1H~ FEKRGI DVDDDASNLKVI LLELA ( SEQ ID NO: 1381 )

[0676] G1 neo2 40 STKKTQLLAEHALLDAYMMLNLVPEPNEKLNRI ITTMQSWI FTGKIDADGAQELAIEVEELEQE 1I~ YEKRGI DVDDYASNLKVI LLELA ( SEQ ID NO: 1382 )

[0677] G1 neo2 40 STKKTQLMAEHALLDAFMMLNVLPEPNEKLNRI ITTMQSWI FTGKIDGDDAQELAKEVEELEQE 1J- LEKRGI DVDDDASNLKVI LLELA ( SEQ ID NO: 1383 )

[0678] G1 neo2 40 STKKTQLLIEHALLDALDMSRNLPEPNEKLSRI ITTMQSWI FTGKIDGDGAQQLAKEVEELEQE 1F~H1 HEKRGEDVEDEASNLKVI LLELA ( SEQ ID NO: 1384 )

[0679] G1 neo2 40 STKKTQLLLEHALLDALHMRRNLPEPNEKLSRI ITTMQSWI FTGKIDGDGAQELAKEVEELEQE 1F~H2 HEKRGRDVEDDASNLKVI LLELA ( SEQ ID NO: 1385 )

[0680] G1 neo2 40 STKKTQLLIEHALLDALNMRKKLPEPNEKLSRI ITDMQSWI FTGKIDGDGAQQLAKEVEELEQE 1F~H3 HEKRGGDVEDYASNLKVI LLELA ( SEQ ID NO: 1386 )

[0681] G1 neo2 40 STKKTQLLLEHALLDALHMSRELPEPNEKLNRI ITDMQSWI FTGKIDGDGAQDLAKEVEELEQE 1F~H4 HEKRGGDVEDYASNLKVI LLELA ( SEQ ID NO: 1387 )

[0682] G1 neo2 40 STKKTQLLIEHALLDALHMSRKLPEPNEKLSRI ITTMQSWI FTGKIDGDGAQHLAKEVEELEQE 1F~H5 HEKRGGEVEDEASNLKVI LLELA ( SEQ ID NO: 1388 )

[0683] G1 neo2 40 STKKTQLLIEHALLDALHMKRKLPEPNEKLNRI ITNMQSWI FTEKIDGDGAQDLAKEVEELEQE 1F~H6 HEKRGQDVEDYASNLKVI LLELA ( SEQ ID NO: 1389 )

[0684]

[0685] G1 neo2 40 STEKTQLAAEHALRDALMLKHLLNEPNEKLARIITTMQSWQFTGKIDGDGAQELAKEVEELQQE 1F~M1 HEVRGI DVEDYASNLKVI LLHLA ( SEQ ID NO: 1390 )

[0686] G1 neo2 40 STKNTQLAAEDALLDALMLRNLLNEPNEKLARIITTMQSWQFTEKIDGDGAQELAKEVEELQQE 1F_M2 HEERGI DVEDYASNLKVI LLQLA ( SEQ ID NO: 1391 )

[0687] G1 neo2 40 STEKTQHAAEDALRDALMLRNLLNEPNEKLARIITTMQSWQFTEKIDGDGAQELAKEVEELQQE 1F~ M3 HEVRGI DVEDYASNLKVI LLQLA ( SEQ ID NO: 1392 )

[0688]

[0689] Table 13C. JL-2 receptor PYc heterodimer binders

[0690] G2 neo2 40 TQKKQQLLAEHALLDALMILNMLKTSSEAVNRMITIAQSWIFTGTSNPEEAKEMIKMAEQAEEE IF seq02 ARREGVDTEDYVSNLKVILKEIA ( SEQ ID NO: 1393 )

[0691] G2 neo2 40 TTKKYQLLVEHALLDALMMLNLSSESNEKMNRI ITTMQSWI FTGTFDPDQAEELAKLVEELREE IF seq03 FRKRGIDTEDYASNLKVILKELS ( SEQ ID NO: 1394 )

[0692] G2 neo2 40 TTKKIQLLVEHALLDALMILNLSSESNEKLNRIITTLQSWTFRGEIDPDRARELAKLLEEIREE IF seq04 MRKRGIDTEDYVSNMIVIIRELA ( SEQ ID NO: 1395 )

[0693] G2 neo2 40 TKKKIQLLAEHVLLDLLMMLNLSSESNEKMNRLITIVQSVJIFTGTIDPDQAEEMAKWVEELREE IF seq05 FRKRGI DTEDYASNVKVI LKELS ( SEQ ID NO: 1396)

[0694] G2 neo2 40 TKKKYQLLIEHLLLDALMVLNMSSESNEKLNRIITILQSWIFTGTWDPDLAEEMEKLMQEIEEE IF seq06 LRRRGIDTEDYMSNMRVIIKELS ( SEQ ID NO: 1397 )

[0695] G2 neo2 40 TKKKLQLLVEHLLLDMLMI LNMSSESNEKLNRLITELQSWI FRGEI DPDKAEEMWKIMEEI EKE IF seq07 LRERGIDTEDYMSNAKVIIKELS ( SEQ ID NO: 1398 )

[0696] G2 neo2 40 TSKKQQLLAEHALLDALMILNISSESSEAVNRAITWLQSWIFKGTVNPDQAEEMRKLAEQIREE lF~seq013 MRKRGIDTEDYVSNLEVI AKELS ( SEQ ID NO: 1399 )

[0697] G2 neo2 40 TKKKYQLLIEHLLLDLLMVLNMSSESNEKINRLITWLQSWIFTGTYDPDLAEEMYKILEELREE IF seq09 MRERGIDTEDYMSNMRVIVKELS ( SEQ ID NO: 1400 )

[0698] G2 neo2 40 TKKKWQLLIEHLLLDLLMILNLSSESNEKLNRLITWLQSWIFTGTYDPDLAEEMKKMMDEIEDE IF seqlO LRERGIDTEDYMSNAKVIIKELS ( SEQ ID NO: 1401 )

[0699] G2 neo2 40 TKKKIQLLVEHALLDALMILNLSSESNEKLNRI ITTMQSWI FTGTIDPDQAEELSKLVEEIREE IF seqll MRKRGIDTEDYVSNLKVILDELS ( SEQ ID NO: 1402 )

[0700] G2 neo2 40 TEKKLQLLVEHALLDALMI LNLWSESNEKLNRI ITTMQS WI FTGRIDPDKAEELAKLVEELREE IF seql2 ARERGIDTEDYVSNLKVILKELS ( SEQ ID NO: 1403 )

[0701] G2 neo2 40 TKKKYQLLMEHLLLDLLWLNMSSESNEKLNRLITIIQSWIFTGTWDPDKAEEMAKMLKEIEDE IF seql3 LRERGIDTEDYMSNMIVIMKELS ( SEQ ID NO: 1404 )

[0702] G2 neo2 40 TTKKIQLLVEHALLDALMLLNLSSESNEKMNRI ITTMQSWI FEGRIDPDQAQELAKLVEELREE IF seql4 FRKRGIDTEDYVSNLKVILEELS ( SEQ ID NO: 1405 )

[0703] G2 neo2 40 T KK K I Q L LVEHAL L DALMMLN L S S E S N E KLNRI ITTMQSWI FT GT I DPDQAEELAKLVRELREE IF seql5 FRKRGI DTEDYASNLEVILRELS ( SEQ ID NO: 1406)

[0704] G2 neo2 40 TKKKIQLLVEHALLDALMILNLSSKSNEKLNRI ITTMQSWI FNGTIDPDRARELAKLVEEIRDE IF seql6 MEKNGIDTEDYVSNLKVI LEELA ( SEQ ID NO: 1407 )

[0705]

[0706] G2 neo2 40 TKKKYQLLIEHVLLDLLMLLNLSSESNEKMNRLITILQSWIFTGTYDPDKAEEMAKLLKELREE IF seql7 FRERGIDTEDYISNAIVILKELS (SEQ ID NO: 1408 )

[0707] G2 neo2 40 TKKKIQLLVEHALLDALMMLNLSSESNEKLNRIITTMQSWIFTGTIDPDRAEELAKLVEELREE lF_seql8 FRKRGIDTEDYASNLKVILKELS (SEQ ID NO: 1409)

[0708] G2 neo2 40 TKKKIQLLVEHALLDALMMLNLSSESNEKLNRIITTMQSWIFNGTIDPDQARELAKLVEELP. EE IF seq!9 FRKRGIDTEDYASNLKVILEELA (SEQ ID NO: 1410)

[0709] G2 neo2 40 TKKKLQLLVEHALLDALMLLNLSSESNEKLNRIITTMQSWIFTGTVDPDQAEELAKLVEEIREE IF seq20 LRKRGIDTEDYVSNLKVILKELS (SEQ ID NO: 1411)

[0710] G2 neo2 40 TTKKYQLLVEHALLDALMILNLSSESNEKLNRIITTMQSWIFTGTFDPDQAEELAKLVREIREE IF seq21 MRKRGIDTEDYVSNLEVILRELS (SEQ ID NO: 1412 )

[0711] G2 neo2 40 TKKKIQLLVEHALLDALMILNLSSESNEKLNRIITTMQSWIFTGTIDPDRAEELAKLVREIP. EE IF seq22 MRKRGIDTEDYVSNLEVILRELS (SEQ ID NO: 1413)

[0712] G2 neo2 40 TKKKYQLLIEHLLLDLLMILNLSSESNEKLNRLITWLQSWIFRGEWDPDKAEEWAKILKEIREE IF seq23 LRERGIDTEDYMSNAIVIMKELS (SEQ ID NO: 1414 )

[0713] G2 neo2 40 TDKKLQLLVEHLLLDLLMMLNLSSKSNEKMNRLITIAQSWIFTGKVDPDLAREMIKLLEETEDE IF seq24 NRKNGIDTEDYVSNARVIAKELE (SEQ ID NO: 1415)

[0714] G2 neo2 40 TKKKIQLLVEHALLDALMLLNLSSESNEKMNRI ITTMQSWI FTGTIDPDQAEELAKLVEELKEE IF seq25 FKKRGIDTEDYVSNLKVILKELS (SEQ ID NO: 1416)

[0715] G2 neo2 40 TKKKYQLLIEHALLDALMI LN LWSESNEKLNRI ITTMQSWI FT GT Y D P D KAE E L E K LAKE I E D E IF seq26 ARERGI DTEDYMSNLRVI LKELS (SEQ ID NO: 1417 )

[0716] G2 neo2 40 TKKKAQLLAEHALLDALMLLNLSSESNERLNRIITWLQSIIFTGTYDPDMVKEAVKLADEIEDE IF seq27 MRKRGIDTEDYVSNLRVILQELA (SEQ ID NO: 1418 )

[0717] G2 neo2 40 TQKKNQLLAEHLLLDALMVLNQSSESSEVANRIITWAQSWI FEGRVDPNKAEEAKKLAKKLEEE IF seq28 MRKRGIDMEDYISNMKVIAEEMS (SEQ ID NO: 1419)

[0718] G2 neo2 40 EDYYSNLKVILEEIAREMERNGLSDKAEEWRQWKKIVERIRQIRSNNSDLNEAKELLNRLITYI IF seq29 QSQIFEISERIRETDQEKKEESWKKWQLLLEHALLDVLMLLND (SEQ ID NO: 1420) G2 neo2 40 PEKKRQLLLEHILLDALMLLNLXXXXXXNTESKFEDYISNAEVIAEELAKLMESXXLSDEAEKF IF seq30 KKIKQWLREVWRIWXXXXWSTLEDKARELLNRIITTIQSQIFY (SEQ ID NO: 1421) PEKKRQLLLEHILLDALMLLNLLETNPQNTESKFEDYISNAEVIAEELAKLMESLGLSDEAEKF KKIKQWLREVWRIWSSTNWSTLEDKARELLNRIITTIQSQIFY (SEQ ID NO: 1422 ) G2 neo2 40 PEKKRQLLLEHILLDLLMILNMXXXXXXNTESEMEDYWSNVRVILRELARLMEEXXXKELSELM IF seq31 ERMRKIVEKIRQIVTXXXXLDTAREWLNRLITWIQSLIFR (SEQ ID NO: 1423)

[0719] PEKKRQLLLEHILLDLLMILNMIETNRENTESEMEDYWSNVRVILRELARLMEELNYKELSELM ERMRKIVEKIRQIVTNNSSLDTAREWLNRLITWIQSLIFR (SEQ ID NO: 1424 ) G2 neo2 40 PEKKRQLLAEHALLDALMLLNIIETNSKNTESKMEDYVSNLEVILTEFKKLAEKLNFSEEAERA IF seq32 ERMKRWARKAYQMMTLDLSLDKAKEMLNRIITILQSIIFN (SEQ ID NO: 1425) G2 neo2 40 PEKKRQLLAEHLLLDVLMMLNGNASLKDYASNAQVIADEFRELARELGLTDEAKKAEKIIEALE IF seq33 RAREWLLNNKDKEKAKEALNRAITIAQSWIFN (SEQ ID NO: 1426)

[0720] G2 neo2 40 PEKKRQLLLEHLLLDLLMILNMLRTNPKNIESDWEDYMSNIEVIIEELRKIMESLGRSEKAKEW IF seq34 KRMKQWVRRILEIVKNNSDLEEAKEWLNRLITIVQSEIFE (SEQ ID NO: 1427 )

[0721]

[0722] G2 neo2 40 WEKKRQLLLEHLLLDLLMILNMWRTNPQNTESLMEDYMSNAKVIVEELARMMRSQGLEDKAREW IF seq35 EEMKKRIEEIRQIIQNNSSKERAKEELNRLITYVQSEIFR ( SEQ ID NO: 1428 ) G2 neo2 40 PKKKIQLLAEHALLDALMILNIVKTNSQNAEEKLEDYASNVEVILEEIARLMESGDQKDEAEKA IF seq36 KRMKEWMKRIKTTASEDEQEEMANRIITLLQSWIFS ( SEQ ID NO: 1429 )

[0723] G2 neo2 40 PEKKRQLLAEHALLDAljMILNXXXXXXQNAEEKLEDYMSNVEVIMEEFARMMRXXXXSEEAENA IF seq37 ERIKKWVRKASSXXXSEEQREMMNRAITLMQSWIFE ( SEQ ID NO: 1430 )

[0724] PEKKRQLLAEHALLDALMILNILQTNPQNAEEKLEDYMSNVEVIMEEFARMMRNGDRSEEAENA ERIKKWVRKASSTASSEEQREMMNRAITLMQSWIFE ( SEQ ID NO: 1431 ) G2 neo2 40 PEKKRQLLAEHLLLDALMVLNMXXXXXXNTEEKLEDYISNMKVIIKEMIELMRSLXXXEEAEKW IF seq38 KEALKAVEKIXXXXDSETARELANRIITLAQSAIFY ( SEQ ID NO: 1432 )

[0725] PEKKRQLLAEHLLLDALMVLNMLTTNSKNTEEKLEDYISNMKVIIKEMIELMRSLGRLEEAEKW KEALKAV'EKIGSRMDSETARELANRIITLAQSAIFY ( SEQ ID NO: 1433 ) G2 neo2 40 PEKKRQLLAEHALLDALMFLNLXXXXXXQAEEKIEDYASNLRVIAEELARLFENLXXXDEAQKA IF seq39 KDIKELAERARSXXSSEKRKEAMNRAITILQSMIFR ( SEQ ID NO: 1434 )

[0726] PEKKRQLLAEHALLDALMFLNLVETNPDQAEEKIEDYASNLRVIAEELARLFENLGRLDEAQKA KDIKELAERARSRVSSEKRKEAMNRAITILQSMIFR ( SEQ ID NO: 1435 ) G2 neo2 40 PEKKRQLLAEHALLDALMILNIIRTNSDNTESKLEDYISNLKVILEEIARLMESLGLSDEAEKA IF seq40 KEAJ4RLADKAGSTASEEEKKEAMNRVITWAQSWIFN ( SEQ ID NO: 1436)

[0727] G2 neo2 40 PEKKRQLLAEHALLDALMT4LNILRTNPDNAEEKLEDYWSNLIVILREIAKLMESLGLTDEAEKA IF seq41 KEAARWAEEARTTASKDQRREIANRIITLLQSWIFS ( SEQ ID NO: 1437 )

[0728] G2 neo2 40 PEKKRQLLAEHLLLDALMILNIIETNEQNAESKLEDYISNAK\riLDEFREMARDLGLLDEAKKA IF seq42 EKMKRWLEKMRSNASSDERREWANRMITTAQSWIFN ( SEQ ID NO: 1438 )

[0729]

[0730] Table 13D. JL-2 receptor PYc heterodimer binders

[0731] G2 neo2 40 TNKKAQLHAEFALHDALMLLNLSSESNERLNRI ITWLQS I I FYGTYDPDMVKEAVKDADEI EDE 1F~ seq27_sJ MRKRGIDTEDYVSNLRLILQELA ( SEQ ID NO: 1439 )

[0732] G2 neo2 40 TNKEAQLHAEFALYDALMLLNLSSESNERLNRI ITWLQS I I FYETYDPDMVKEAVKLADEI EDE 1F~ seq27_sT MRKRKIDTEDYWNLRLILQELA ( SEQ ID NO: 1440 )

[0733] 8

[0734] G2 neo2 40 TKKDAELLAEFALYDALMLLNLSSESNERLNEIITWLQSIIFYGTYDPDMVKEAVKLADEIEDE 1F~ seq27_slE MRKRGIDTEDYVSNLRLILQELA ( SEQ ID NO: 1441 )

[0735] 2

[0736] G2 neo2 40 TNKKAQLHAEFALYDALMLLNLSSESNERLNDIITWLQSIIFTGTYDPDMVKEAVKLADEIEDE 1F~ seq27_slE MRKRKIDTEDYWNLRYILQELA ( SEQ ID NO: 1442 )

[0737] 4

[0738] G2 neo2 40 EDYYSNLKLILEELAREMERNGLSDKAEEWRQWKKIVERIRQIRSNNSDLNEAKELLNRLITYI IF seq29 S6 QSQIFEVLHGVGETDQEKKEESWKKWDLLLEHALLDVLMLLND ( SEQ ID NO: 1443 ) G2 neo2 40 EDYYSNLKVILEELAREMERNGLSDKAEEWRQWKKIVERIRQIRSNNSDLNEAKELLNELITYI IF seq29 S7 QSQIFEVIEREGETDQEKKEESWKKWELHLEHALLDVLMLLND ( SEQ ID NO: 1444 ) G2 neo2 40 EDYYSNLKLILEELAREMERNGLSDKAEEWRQWKKIVERIRQIRSNNSDLNEAKELLNRLITYI 1F~ seq29_sl3 QSQIFEVLEGVGETDQEKKEESWKKWELHLEHALLDVLMLLND ( SEQ ID NO: 1445 ) Neoleukin- PKKKIQLHAEHALYDALMILNIVKTNSPPAEEKLEDYAFNFELILEEIARLFESGDQKDEAEKA

[0739]

[0740] 2 / 15 KRMKEWMKRIKTTASEDEQEEMANAIITILQSWIFS ( SEQ ID NO: 1446)(i. e.

[0741] G2 neo2 40

[0742] IF seq36 SI

[0743] 1 )“

[0744] G2 neo2 40 PKKKIQLLAEHALFDLLMILNIVKTNSQNAEEKLEDYAYNAGVILEEIARLFESGDQKDEAEKA IF seq36 SI KRMKEWMKRIKDTASEDEQEEMANEIITILQSWNFS ( SEQ ID NO: 1447 )

[0745] 2

[0746] Neoleukin- PKKKIQLYAEHALYDALMILNIVKTNSPPAEEELEDYAFNFELILEEIARLFESGDQKDEAEKA 2 / 15-H8Y- KRMKEWMKRIKTTASEDEQEEMANAIITILQSWIFS ( SEQ ID NO: 1448 )

[0747] K33E

[0748]

[0749] Table 14. Other binders (residues in parentheses are optional)

[0750] act 10 4 TEHVARFTALAALEMAKYRRDPEDMRHFLELAERAAHEANDPELEEEVERVKEEL ( SEQ ID NO: 1503

[0751] ActRIIa

[0752] binder

[0753] act 4 7 15 PEKEVAELFHKAREAFQKRDIEKLKEIEKKIEELYKETGQPAVRHPLELVKFFIEELE ( SEQ ID NO: 1504 )

[0754] ActRIIa

[0755] binder

[0756] actriib bp NRWRATARFAMRMAKIFIERGNKERARHFLDQAMRIAELQGDEELLKEVRELLEELE SEQ ID NO: 1505 )

[0757] ActRIIB

[0758] binder

[0759] Alk2 bp DEKRRQALEEIIAAGKEEGVTGEIHIKKGVFEVKIHAENQEKRDAVEERIKEIAAKHGLKVNIL P ( SEQ ID NO: 1506)

[0760] Alk3 bp DYTLTKTGHLENGKGTLTLTLTSPDGKVATATVPMELDKETGAFVLPREKFLAALKELKAQLEA

[0761]

[0762] (K) ( SEQ ID NO: 1507 )

[0763] Tablel5 (Residues in parentheses are optional)

[0764] Design SEQ Hetero N-term C-term Sequence

[0765] name in ID fusion mb SEQ mb SEQ

[0766] example NO: ID NO ID NO

[0767] s

[0768] Alklmb_ 1449 Alkl_H 37 1502 (MSGG) QEEIFRALALFSADLLNIEDVKIE Her2mb er2 SKDGRVKVTVKGNSPDSEEFERYLRELAER LGLEVEIERTGGSGGSGGSGSVDNKFNKEM RNAYWEIALLPNLNNQQKRAFIRSLYDDPS QSANLLAEAKKLNDAQAP (GSGSHHWGSTH HHHHH) *

[0769] Alklmb_ 1450 Alkl_I 37 45 (MSGG) QEEIFRALALFSADLLNIEDVKIE Lytacmb GF_End SKDGRVKVTVKGNSPDSEEFERYLRELAER oTag4 LGLEVEIERTGGSGGSGGSGMEEAQRLLLE WEAQEWADSQGEDAKRVKQWQQILQDPDL EVQKKMLEILKRIYEEK (GSGSHHWGSTHH HHHH) *

[0770] Alklmb_ 1451 Alkl_E 37 1357 (MSGG) QEEIFRALALFSADLLNIEDVKIE EGFRcmb GFRc SKDGRVKVTVKGNSPDSEEFERYLRELAER

[0771] LGLEVEIERTGGSGGSGGSGSLDEAKKLLQ

[0772]

[0773] EAEKLARKLNDRTELAYVEFLKHSLETAKKQNDKRTIESVRDMARDALEELQS (GSGSHH WGSTHHHHHH) *

[0774] Alklmb_ 1452 Alkl_G 37 1358, (MSGG) QEEIFRALALFSADLLNIEDVKIE Gamma Cm ammaC 1359 SKDGRVKVTVKGNSPDSEEFERYLRELAER b LGLEVEIERTGGSGGSGGSGSDVEEVFLRS ARELIKAIERTGDPELEENLQQAMWALRGI KSEEAEELLERAEKLL (GSGSHHWGSTHHH HHH) *

[0775] Alklmb_ 1453 Alkl_I 37 111 (MSGG) QEEIFRALALFSADLLNIEDVKIE Insulin nsulin SKDGRVKVTVKGNSPDSEEFERYLRELAER Rmb R LGLEVEIERTGGSGGSGGSGSKLEEIEELL KELSKTNPLAKDILWVIEVRTEDGHDPKSE LVFIRQYLKTLNTPEAREILKIVAP (GSGS HHWGSTHHHHHH) *

[0776] EGFRcmb 1454 EGFRc_ 1357 39 (MSG) SLDEAKKLLQEAEKLARKLNDRTEL _TFGBR2 TFGBR2 AYVEFLKHSLETAKKQNDKRTIESVRDMAR mb DALEELQSGGSGGSGGSGGLKELLKELNKA IASGDTETVRRILEELLELLKEAFEKGDYD LAISIASMAVKAASYIGDTETLKELLEILK KI KEKLKKEGDEAALKAVERNI KWEKVA ( GSGSHHWGSTHHHHHH) *

[0777] Gamma Cm 1456 Gamma C 1358, 1335 (MSG ( SDVEEVFLRSARELIKAIERTGDPE b_PDGFR _PDGFR 1359 LEENLQQAMWALRGIKSEEAEELLERAEKL mb LGGSGGSGGSGSEQDKQVKKLTDIQREAYK HNDKETIEKALRTAASFARQHRDPKFRELF RIIHQLRQRVER (GSGSHHWGSTHHHHHH) Her2af f 1457 Her2_F 1502 11, 1031 (MSG) SVDNKFNKEMRNAYWEIALLPNLNN _FGFRmb GFR QQKRAFIRSLYDDPSQSANLLAEAKKLNDA QAPGGSGGSGGSGDRRKEMDKVYRTAYKRI TSTPDKEKRKEWKEATEQLRRIAKDEEEK KKAAYMISFLKTLG (GSGSHHWGSTHHHHH H) *

[0778] IGF2Rmb 1458 IGF2R_ 40 1335 (MSG) TERRVIQVLEQILEDEDPEWEKML _PDGFRm PDGFR EILLEILEEAGDPARKLVEEILRWRKNLE b EARELVRRLSGGSGGSGGSGSEQDKQVKKL TDIQREAYKHNDKETIEKALRTAASFARQH RDPKFRELFRIIHQLRQRVER (GSGSHHWG STHHHHHH) *

[0779] Insulin 1459 Insuli 111 1358, (MSG) SKLEEIEELLKELSKTNPLAKDILW Rmb Gam nR Gam 1359 VIEVRTEDGHDPKSELVFIRQYLKTLNTPE maCmb maC AREILKIVAPGGSGGSGGSGSDVEEVFLRS ARELIKAIERTGDPELEENLQQAMWALRGI KSEEAEELLERAEKLL (GSGSHHWGSTHHH HHH) *

[0780] Lytacmb 1460 IGF_En. 37 (MSG) MEEAQRLLLEWEAQEWADSQGEDAK _ALKlmb doTag4 RVKQWQQILQDPDLEVQKKMLEILKRIYE _ALK1 EKGGSGGSGGSGGQEEIFRALALFSADLLN IEDVKIESKDGRVKVTVKGNSPDSEEFERY LRELAERLGLEVEIERT ( GSGSHHWGSTHH HHHH) *

[0781] TGFBR2m 1461 TGFBR2 39 1033 (MSGG) LKELLKELNKAIASGDTETVRRIL b Gp 130 _Gpl30 EELLELLKEAFEKGDYDLAI S I ASMAVKAA mb SYIGDTETLKELLEILKKIKEKLKKEGDEA ALKAVERNIKWEKVAGGSGGSGGSGDISE RFRRLMRRAD E LARRGN P E EARKVL E EAE E LMERYGSPELLESVRMLLEVLG (GSGSHHW GSTHHHHHH) *

[0782] TGFBR2m 1462 TGFBR2 39 39 (MSGG) LKELLKELNKAIASGDTETVRRIL b_TGFBR _TGFBR EELLELLKEAFEKGDYDLAI S I ASMAVKAA

[0783]

[0784] 2mb 2 SYIGDTETLKELLEILKKIKEKLKKEGDEAALKAVERNIKWEKVAGGSGGSGGSGGLKE LLKELNKAIASGDTETVRRILEELLELLKE AFEKGDYDLAISIASMAVKAASYIGDTETL KELLEILKKIKEKLKKEGDEAALKAVERNI KWEKVA (GSGSHHWGSTHHHHHH) * TGFBR2m 1463 TGFBR2 39 (MSGG) LKELLKELNKAIASGDTETVRRIL b Her2m Her2 EELLELLKEAFEKGDYDLAI S I ASMAVKAA b~ SYIGDTETLKELLEILKKIKEKLKKEGDEA ALKAVERNIKWEKVAGGSGGSGGSGSVDN KFNKEMRNAYWEIALLPNLNNQQKRAFIRS LYDDPSQSANLLAEAKKLNDAQAP (GSGSH HWGSTHHHHHH) *

[0785] TrkAmb_ 1464 TrkA_B 16, 28 31 (MSG) RDEIKERIKKAWRARVTGNPEQLK BMPR2mb MPR2 EAKKLLEKLKKNGRDDQDAKKFEKAI RQVE KRLRSGGSGGSGGSGTEEEKVKKLIEKIRE AAKRGDRHLRYRLLHELERI AVKLGDWRI L VQLVEAAKEAEEIN (GSGSHHWGSTHHHHH H) *

[0786] TGFbR2m 1465 TGFbR2 39 16, 28 (MSGG) LKELLKELNKAIASGDTETVRRIL b_TrkAm _TrkA EELLELLKEAFEKGDYDLAI S I ASMAVKAA b SYIGDTETLKELLEILKKIKEKLKKEGDEA ALKAVERNIKWEKVAGGSGGSGGSGRDEI KERIKKAWRARVTGNPEQLKEAKKLLEKL KKNGRDDQDAKKFEKAIRQVEKRLRS (GSG SHHWGSTHHHHHH) *

[0787] TGFbR2m 1466 TGFbR2 39 31 (MSGG) LKELLKELNKAIASGDTETVRRIL b_BMPR2 _BMPR2 EELLELLKEAFEKGDYDLAI S I ASMAVKAA mb SYIGDTETLKELLEILKKIKEKLKKEGDEA ALKAVERNIKWEKVAGGSGGSGGSGTEEE KVKKLI EKI REAAKRGDRHLRYRLLHELER IAVKLGDWRILVQLVEAAKEAEEIN (GSGS HHWGSTHHHHHH) *

[0788] Alklmb_ 1467 Alkl_A 37 (MSGG) QEEIFRALALFSADLLNIEDVKIE Alklmb Ikl SKDGRVKVTVKGNSPDSEEFERYLRELAER LGLEVEIERTGGSGGSGGSGGQEEIFRALA LFSADLLNIEDVKIESKDGRVKVTVKGNSP DSEEFERYLRELAERLGLEVEIERT (GSGS HHWGSTHHHHHH) *

[0789] Alkl_IG 1468 Alkl_I 37 40 (MSGG) QEEIFRALALFSADLLNIEDVKIE F2R GF2R SKDGRVKVTVKGNSPDSEEFERYLRELAER LGLEVEIERTGGSGGSGGSGTERRVIQVLE QILEDEDPEWEKMLEILLEILEEAGDPAR KLVEEILRWRKNLEEARELVRRLS (GSGS HHWGSTHHHHHH) *

[0790] Gamma Cm 1469 Gamma C 1358, 39 (MSG) SDVEEVFLRSARELIKAIERTGDPE b_TGFBR _TGFBR 1359 LEENLQQAMWALRGIKSEEAEELLERAEKL 2mb 2 LGGSGGSGGSGGLKELLKELNKAIASGDTE TVRRI LEELLELLKEAFEKGDYDLAI S IAS MAVKAASYIGDTETLKELLEILKKIKEKLK KEGDEAALKAVERNIKWEKVA ( GSGSHHW GSTHHHHHH) *

[0791] Her2af f 1470 Her2_T 1502 39 (MSG) SVDNKFNKEMRNAYWEIALLPNLNN _TGFBR2 GFBR2 QQKRAFIRSLYDDPSQSANLLAEAKKLNDA mb QAPGGSGGSGGSGGLKELLKELNKAIASGD TETVRRI LEELLELLKEAFEKGDYDLAI SI ASMAVKAASYIGDTETLKELLEILKKIKEK LKKEGDEAALKAVERNIKWEKVA ( GSGSH HWGSTHHHHHH) *

[0792] Neo2mb 1472 Neo2_E 1360, 1357 (MSGG) SHMPKKKIQLHAEHALYDALMILN EGFRcmb GFRc 1361, IVKTNSPPAEEKLEDYAFNFELILEEIARL

[0793]

[0794] 1446 FESGDQKDEAEKAKRMKEWMKRIKTTASEDEQEEMANAIITILQSWIFSGGSGGSGGSGS LDEAKKLLQEAEKLARKLNDRTELAYVEFL KHSLETAKKQNDKRTIESVRDMARDALEEL QS (GSGSHHWGSTHHHHHH) * TGFBR2m 1472 TGFBR2 S9 40 (MSGG) LKELLKELNKAIASGDTETVRRIL b_IGF2R _IGF2R EELLELLKEAFEKGDYDLAI S I ASMAVKAA mb SYIGDTETLKELLEILKKIKEKLKKEGDEA ALKAVERNIKWEKVAGGSGGSGGSGTERR VIQVLEQILEDEDPEWEKMLEILLEILEE AGD P ARKL VE E I L RWRKN L E EARE L VRRL

[0795] S (GSGSHHWGSTHHHHHH) * Her2mb 1474 Her2mb 2 11 (MSG) SVDEKIEELYEKVKELAKKGDREAT FGFR _FGFR AKVLAELYRLAVQSGDDKVFDRLDEAYQTA (H2F) RENLSGSGSGSGSGSDRRKEMDKVYRTAYK RITSTPDKEKRKEWKEATEQLRRIAKDEE EKKKAAYMISFLKTLG ( GSGSHHWGSTHHH HHH) *

[0796] FGFR_He 1475 FGFR_H 2 11, 10S1 (MSG) DRRKEMDKVYRTAYKRITSTPDKEK r2mb er2mb RKEWKEATEQLRRIAKDEEEKKKAAYMI S ( FH2 ) FLKTLGGSGSGSGSGSSVDEKIEELYEKVK ELAKKGDREATAKVLAELYRLAVQSGDDKV FDRLDEAYQTARENLS (GSGSHHWGSTHHH HHH) *

[0797] BMPR2mb 1479 BMPR2_ SI 2 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR Her2 m Her2 m YRLLHELERI AVKLGDWRI LVQLVEAAKEA b b EEINGGSGGSGGSGSVDEKIEELYEKVKEL AKKGDREATAKVLAELYRLAVQSGDDKVFD RLDEAYQTARENLS ( GSGSHHWGSTHHHHH H) *

[0798] act 10 1480 act 10 150311, 10S1 (MSG) TEHVARFTALAALEMAKYRRDPEDM 4_FGFR—_4_FGF RHFLELAERAAHEANDPELEEEVERVKEEL R GSGSGSGDRRKEMDKVYRTAYKRITSTPDK EKRKEWKEATEQLRRIAKDEEEKKKAAYM ISFLKTLG (GSGSHHWGSTHHHHHH) * act 10 1481 act 10 15031S57 (MSG) TEHVARFTALAALEMAKYRRDPEDM 4_EGFRc _4_EGF RHFLELAERAAHEANDPELEEEVERVKEEL Rc GSGSGSGSLDEAKKLLQEAEKLARKLNDRT ELAYVEFLKHSLETAKKQNDKRTIESVRDM ARDALEELQS (GSGSHHWGSTHHHHHH) * act 10 1482 act 10 150316, 28 (MSG) TEHVARFTALAALEMAKYRRDPEDM 4_TrkA _4_Trk RHFLELAERAAHEANDPELEEEVERVKEEL A GSGSGSGRDEIKERIKKAWRARVTGNPEQ LKEAKKLLEKLKKNGRDDQDAKKFEKAI RQ VEKRLRS (GSGSHHWGSTHHHHHH) * BR2_2_31482 BR2_2_ SI 1S57 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR _EGFRc3_EGFR YRLLHELERI AVKLGDWRI LVQLVEAAKEA c EEINGSGSGSGSLDEAKKLLQEAEKLARKL NDRTELAYVEFLKHSLETAKKQNDKRTIES VRDMARDALEELQS (GSGSHHWGSTHHHHH H) *

[0799] BR2_2_31484BR2_2_ SI 1508 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR _IGF1R3_IGF1 YRLLHELERI AVKLGDWRI LVQLVEAAKEA R EEINGSGSGSGSVEERLEEIARKYLPPEDV EFLKLALRALIKDLNVPPERALKQLIWIAR QSGDPELVRVLELALK ( GSGSHHWGSTHHH HHH) *

[0800] TrkA_bm 1485 TrkA_b 16, 2832(MSG) RDEIKERIKKAWRARVTGNPEQLK pr2_l_l mpr2_l EAKKLLEKLKKNGRDDQDAKKFEKAI RQVE m2 1 m2 KRLRSGGSGGSGGSNLEALIFFNRVQARFL

[0801] GSERYLEVANQAEEALRRGDREKAYQILLE

[0802]

[0803] SEKSL (GSGSHHWGSTHHHHHH) *act 10 1486 act 10 1503 1503 (MSG) TEHVARFTALAALEMAKYRRDPEDM 4 act 1 4 act RHFLELAERAAHEANDPELEEEVERVKEEL °24l!0_4 GSGSGSTEHVARFTALAALEMAKYRRDPED MRHFLELAERAAHEANDPELEEEVERVKEE

[0804] L (GSGSHHWGSTHHHHHH) *BR2_2_31487BR2_2_ 31 16, 28 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR _TrkA 3_TrkA YRLLHELERI AVKLGDWRI LVQLVEAAKEA EEINGSGSGSGRDEIKERIKKAWRARVTG NPEQLKEAKKLLEKLKKNGRDDQDAKKFEK AIRQVEKRLRS (GSGSHHWGSTHHHHHH) * alkl vc 1488 alkl v 37 31 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER _cys_BM no cys LGLEVEIERTGSGSGSGTEEEKVKKLIEKI PR2 _BMPR2 REAAKRGDRHLRYRLLHELERIAVKLGDWR ILVQLVEAAKEAEEIN (GSGSHHWGSTHHH HHH) *

[0805] alkl vc 1489 alkl v 37 1506 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER cys Al no cys LGLEVEIERTGSGSGSDEKRRQALEEIIAA k2 bp Alk2 GKEEGVTGEIHIKKGVFEVKIHAENQEKRD bp AVEERIKEIAAKHGLKVNILP (GSGSHHWG STHHHHHH) *

[0806] alkl vc 1490 alkl v 37 1507 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER cys Al no cys LGLEVEIERTGSGSGSDYTLTKTGHLENGK k3 bp Alk3 GTLTLTLTSPDGKVATATVPMELDKETGAF bp VLPREKFLAALKELKAQLEA ( GSGSHHWGS THHHHHH) *

[0807] actriib 1491 actrii 1505 37 (MSG) NRWRATARFAMRMAKIFIERGNKER bp alk b bp a ARHFLDQAMRIAELQGDEELLKEVRELLEE 1 vc3 6 lkl_vc LEGSGSGSGQEEIFRALALFSADLLNIEDV 4 no cy 3_64_n KIESKDGRVKVTVKGNSPDSEEFERYLREL s o cys AERLGLEVEIERT (GSGSHHWGSTHHHHHH ) *

[0808] Alk3 bp 1492 Alk3_b 1507 2 (MSG) DYTLTKTGHLENGKGTLTLTLTSPD Her2 m p Her2 GKVATATVPMELDKETGAFVLPREKFLAAL b mb KELKAQLEAGSGSGSGSVDEKIEELYEKVK ELAKKGDREATAKVLAELYRLAVQSGDDKV FDRLDEAYQTARENLS (GSGSHHWGSTHHH HHH) *

[0809] BR2_2_3 1493BR2_2_ 31 31 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR _BR2_2_ 3_BR2_ YRLLHELERI AVKLGDWRI LVQLVEAAKEA 3 2_3 EEINGSGSGSTEEEKVKKLIEKIREAAKRG DRHLRYRLLHELERI AVKLGDWRI LVQLVE AAKEAEEIN (GSGSHHWGSTHHHHHH) * alkl vc 1494 alkl v 37 39 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER _cys_TG no cys LGLEVEIERTGSGSGSGGLKELLKELNKAI FbR2 _TGFbR ASGDTETVRRILEELLELLKEAFEKGDYDL 2 AISIASMAVKAASYIGDTETLKELLEILKK I KEKLKKEGDEAALKAVERNI KWEKVA ( G SGSHHWGSTHHHHHH) *

[0810] alkl vc 1495 alkl v 37 31 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER _cys_BR no cys LGLEVEIERTGSGSGSTEEEKVKKLIEKIR 2_2_3 _BR2_2 EAAKRGDRHLRYRLLHELERIAVKLGDWRI _3 LVQLVEAAKEAEEIN (GSGSHHWGSTHHHH HH) *

[0811] EGFRc_B 1496 EGFRc_ 1357 31 (MSG) SLDEAKKLLQEAEKLARKLNDRTEL R2_2_3BR2_2_ AYVEFLKHSLETAKKQNDKRTIESVRDMAR

[0812]

[0813] 3 DALEELQSGGSGGSGGSTEEEKVKKLIEKIREAAKRGDRHLRYRLLHELERIAVKLGDWR ILVQLVEAAKEAEEIN (GSGSHHWGSTHHH HHH) *

[0814] Her2 mb 1497 Her2 m 2 31 (MSG) SVDEKIEELYEKVKELAKKGDREAT _BR2~2_ b_BR2_ AKVLAELYRLAVQSGDDKVFDRLDEAYQTA 3 2_3 RENLSGGSGGSGGSTEEEKVKKLIEKIREA AKRGDRHLRYRLLHELERI AVKLGDWRI LV QLVEAAKEAEEIN (GSGSHHWGSTHHHHHH ) *

[0815] BMPR2_B 1498 BMPR2_ 31 31 (MSG) TEEEKVKKLIEKIREAAKRGDRHLR R2_2_3 BR2_2_ YRLLHELERI AVKLGDWRI LVQLVEAAKEA 3 EEINGGSGGSGGSTEEEKVKKLIEKIREAA KRGDRHLRYRLLHELERI AVKLGDWRI LVQ LVEAAKEAEEIN (GSGSHHWGSTHHHHHH) act 10 1499 act 10 1503 1505 (MSG) TEHVARFTALAALEMAKYRRDPEDM 4 actri 4 act RHFLELAERAAHEANDPELEEEVERVKEEL ib bp riibjo GSGSGSNRWRATARFAMRMAKIFIERGNKE P RARHFLDQAMRIAELQGDEELLKEVRELLE ELE (GSGSHHWGSTHHHHHH) *

[0816] Alk3 bp 1500 Alk3_b 1507 32 (MSG) DYTLTKTGHLENGKGTLTLTLTSPD bmpr2 p bmpr GKVATATVPMELDKETGAFVLPREKFLAAL l_l_m2- KELKAQLEAGSGSGSNLEALIFFNRVQARF m2 LGSERYLEVANQAEEALRRGDREKAYQILL ESEKSL (GSGSHHWGSTHHHHHH) * alkl vc 1494 alkl v 37 39 (MSGG) QEEIFRALALFSADLLNIEDVKIE 3 64 no c3_64_ SKDGRVKVTVKGNSPDSEEFERYLRELAER _cys_TG no cys LGLEVEIERTGSGSGSGGLKELLKELNKAI FbR2 _TGFbR ASGDTETVRRILEELLELLKEAFEKGDYDL 2 AISIASMAVKAASYIGDTETLKELLEILKK I KEKLKKEGDEAALKAVERNI KWEKVA ( G

[0817]

[0818] SGSHHWGSTHHHHHH) *

[0819] Table 16

[0820] SEQ ID Designed protein cocktail components sequence

[0821] NO:

[0822] 1476 06- (MG) S S D E E EARE LEE RAREAAKRAI EAAKRT GD P RVRE LAE E L VKLAI WAAVE V 79C_mb7 W L D P S S S DVN EAL KL I VEAI EAAVRAL EAAE RT GD P E VRE LARE L VRLAVEAAE E VQ RN P S S S DVN EAL KL I VI Al EAAVRAL EAAE RT GD P E VRE LARE L VRLAVEAAE E VQ RN P S S E EVN EAL RKIIKLIL FAVMVL E LAE E I GD P T WREMARRAVREAVE LA E E VQ RDPSGWLGH G SDRRKEMDKVYRTAYKRI TS TPDKEKRKEWKEATEQLRRI AKDEEEKKKAAYMISFLKTLGS (LEHHHHHH)

[0823] 37 ALK1 GQEEIFRALALFSADLLNIEDVKIESKDGRVKVTVKGNSPDSEEFERYLRELAER LGLEVEIERT

[0824] 1477 TGFBR2 SGSGLKELLKELNKAIASGDTETVRRILEELLELLKEAFEKGDYDLAISIASMAV ( related KAASYIGDTETLKELLEILKKI KEKLKKEGDEAALKAVERNI KWEKVA to SEQ

[0825] ID

[0826]

[0827] NO: 39 )

[0828] Example 1

[0829] Abstract

[0830] Growth factor induced receptor dimerization and activation of downstream pathways can modulate cell fate decisions. Here, we investigate the potential of de novo designedsynthetic ligands, termed novokines, to reprogram cell identity by inducing proximity of novel pairs of receptor subunits. We find that a design, H2F, that brings together HER2 (which has no known natural ligand) and the FGF receptor has potent signaling activity. H2F induces robust signaling and reprograms fibroblasts into myogenic cells. Unlike native FGF ligands, H2F selectively activates the MAPK pathway without engaging PLCy-mediated Ca2+signaling. FRET assays confirm H2F-mediated HER2-FGFR proximity, and phosphoproteomic analysis reveals activation of MAPK effectors. H2F-induced ERK phosphorylation is abolished in cells expressing a kinase-dead FGFR1 (K514M) mutant, confirming the requirement for FGFR catalytic activity. H2F treatment significantly increases myofiber formation from adult patient-derived primary myoblasts, demonstrating its capacity to promote myogenic regeneration. Our findings demonstrate that synthetic receptor pairings can rewire signaling outputs to drive regeneration, providing a programmable platform for cell fate engineering.

[0831] Introduction

[0832] The direct conversion, or transdifferentiation, of somatic non-muscle cells into skeletal myocytes offers substantial therapeutic potential in the future for treating conditions such as muscle atrophy and sarcopenia. Cell fate transitions are often orchestrated by growth factors that induce the assembly of specific receptor pairs, leading to the transactivation of intracellular kinase domains and the initiation of lineage-specific signaling cascades.

[0833] However, natural growth factors have not been shown to induce fibroblast-to-myoblast conversion, suggesting that new strategies are needed to unlock alternative cell identities. Advances in de novo protein design have enabled the generation of compact (50-110 amino acids), high-affinity receptor-binding domains with remarkable isoform specificity, thermal stability, and modular architecture. These domains typically act as antagonists in monomeric form but can function as potent agonists when presented in multivalent configurations that promote receptor clustering1’2,3. We reasoned that the unique modularity and precision of these synthetic ligands could be leveraged to reprogram cell fate. We investigated whether bringing together receptor subunits that are not known to naturally dimerize, such as HER2 with an unrelated Receptor Tyrosine Kinase (RTK), could trigger noncanonical signaling events capable of driving lineage conversion.

[0834] In this study, utilizing the skeletal muscle transdifferentiation model we identified the HER2: FGFR novokine (H2F) as a potent driver of myogenesis. H2F promotes HER2-FGFR heterodimerization, confirmed by FRET. H2F signaling relies on FGFR kinase activity andY653 / Y654 phosphorylation, with phosphoproteomics revealing activation of MAPK effectors and regulators such as GAPs and GEFs. H2F stands as an exceptional ligand that can bifurcates FGFR pathways where MAPK / AKT activation and eliminates PLCy / Ca2+signaling. Functionally, H2F enhances direct reprogramming of fibroblast into skeletal muscle cells, increases myotube formation from patient-derived myoblasts, and also supports iPSC-derived myogenesis. Like FGF2, H2F preserves iPSC pluripotency, indicating PLCy signaling is dispensable for sternness. But unlike FGF2, H2F does not support endothelial differentiation, highlighting the relevance of Ca2+signaling arm in endothelial cell fate determination. H2F activity could be exploited in various physiological niches where HER2 and FGFRl / 2c are co-expressed. Broadly, our modular strategy of fusing designed binders to recruit unrelated endogenous receptor pairs establishes novokines as reprogramming tools via rewiring novel signaling outputs. Novokines represent a powerful framework for both therapeutic development in regenerative medicine and discovery of fundamentals of signaling logics through receptor remodeling.

[0835] Results

[0836] We screened a set of de novo designed novokines constructed by fusing two computationally designed receptor-binding domains via a flexible linker (Fig. 1 A-C) for effects on skeletal muscle reprogramming. These binding domains are engineered to target a diverse set of Receptor Tyrosine Kinases and Receptor Serine / Threonine Kinases (RTKs / RSTKs) with high affinity and specificity1-3. By simultaneously engaging two distinct receptor subunits, novokines bring these receptors into close proximity. This enforced proximity may promote cross-phosphorylation of intracellular domains or associated signaling components, thereby initiating novel downstream signaling events with the potential to influence cell fate reprogramming

[0837] Designed novokines enhanced skeletal muscle reprogramming

[0838] We began by developing a screen for compounds that increased the reprogramming efficiency of fibroblasts expressing MyoD. MyoD induces direct reprogramming of human fibroblasts into skeletal muscle4’5, but the efficiency of the conversion process remains low6,7,8. Accordingly, even prolonged MyoD overexpression in HFF-iMYOD (a Doxycycline-inducible MyoD overexpression line in human foreskin fibroblasts, Fig. 2A) for 7 or 14 days did not show improvement in full or partial commitment to myogenic fate with the majority of cells exiting the reprogramming pathway (Fig. 2B-C). We optimized the assayfor screening the library of novokines (Fig. 2). The expression of several muscle markers, including MYH3, MYH7, MYH8, TTN3, ACTN2, DESMIN, TITIN, ACT Al (skeletal musclespecific actin), shows that fibroblast fate has partially changed towards trans differentiated myotubes (tSKM) (Fig. 2E). We tested natural ligands (FGF2, FGF7, FGF10, IL-6, EGF, BMP4, VEGF, TNF-a) but did not observe significantly enhanced myogenic conversion (Fig.

[0839] 3D), highlighting the need for synthetic approaches. We therefore screened 155 designed novokines, targeting various combinations of RTKs / RSTKs, added on days 4-8 post-MyoD induction (Fig. 2D), to identify any that enhance myogenic efficiency.

[0840] We identified novokines that passed both primary and secondary screens, significantly enhancing myogenic conversion efficiency, as assessed by Desmin and MHC (Myosin Heavy Chain) expression (Fig. 2D, 2G, 2L; see novokine sequences tested in Table 15 as well as SEQ ID NOS: 12-15 and 33-36).

[0841] To further investigate their mechanisms, we analyzed selected candidate novokines via flow cytometry -based phospho-effectors (pJNK, pP38, pAKT, etc) analysis (Fig. 2E). In starved immortalized EA.hy926 cells (an immortalized human umbilical vein endothelial cell line), a brief 15-minute novokine treatment was sufficient to elicit phospho-effector signaling responses (Fig. 2F).

[0842] We next evaluated the novokine library in an independent myogenic model using induced pluripotent stem cell (iPSC)-derived skeletal muscle differentiation. To this end, we generated a WTC-11 iMYOD cell line, in which MyoD expression is driven by a doxycycline-inducible promoter6(Fig. 21). To assess the ability of novokines to enhance myogenic differentiation, we established an iPSC-derived skeletal myocyte (iSKM) assay: MyoD was induced for six days to initiate myogenic programming, followed by four additional days of differentiation in the presence of individual novokines. We screened the novokine library for effects on differentiation, using desmin and myosin heavy chain (MHC) expression as molecular markers, along with morphological assessments (Fig. 21). A subset of novokines significantly increased desmin and MHC levels (Fig. 21). Among these, 13 novokines also induced hallmark morphological features of myogenic differentiation, including elongated, multinucleated myotubes (Fig. 2J-K). These novokines are:

[0843] ALKI Lytac;

[0844] HER2 TGFBR2;

[0845] ALKl_Gpl30;

[0846] IGF1R PDGFR;BMPR2 TRKA;

[0847] PDGFR Lytac;

[0848] PDGFR ALKl;

[0849] EGFRc PDFGR

[0850] EGFRcside lnsulinR; and

[0851] BMPR2 IGF2R.

[0852] FGFRmb: HER2mb

[0853] EGFRc si demb: BM PRI Imb

[0854] ALKlmb: TRKAmb

[0855] In this study, we focus on a specific novokine, HER2aff-FGFRmb (H2affF) (SEQ ID NO: 1457), which promoted myogenic differentiation in both the fibroblast transdifferentiation assay and the iPSC-derived skeletal muscle model. H2F comprises an FGFR-binding minibinder (FGFRmb) fused to a HER2-specific affibody (H2aff)1. To independently test whether bringing HER2 and FGFR into proximity is sufficient to trigger signaling, we engineered a new minibinder for HER2. HER2, a member of the EGF receptor family of receptor tyrosine kinases (RTKs), is known to heterodimerize with EGFR upon ligand binding to initiate downstream signaling9. A HER2 minibinder (H2mb) that binds to the extracellular domain IV of HER2 distinct from the affibody binding site (Fig. 1C) using Rifgen (Cao et al), was designed followed by ProteinMPNN™ and AlphaFold™ filtering as outlined by Bennett et al10. Candidates were screened via yeast surface display (YSD) and ranked by binding affinity. The top-performing binder was optimized through site- saturation mutagenesis (SSM), and enriched variants were incorporated into a degenerate codon combinatorial library, which was again screened by YSD3. 96 top variants were assessed for soluble expression in ’. coli and binding affinity via biolayer interferometry (BLI / Octet™) (Fig.1C). A single lead candidate with robust soluble expression and sub-nanomolar binding affinity was selected for fusion with FGFRmb and subsequent functional characterization.

[0856] HER2mb: FGFRl / 2cmb mediated receptor heterodimerization augments muscle Reprogramming

[0857] To gain molecular insights into HER2 and FGFRl / 2c heterodimerization, we expressed three constructs- HER2mb-FGFRmb (H2F) (SEQ ID NO: 14-15), FGFRmb-HER2mb (FH2) (SEQ ID NO: 12-13), and HER2aff-FGFRmb (H2affF) (SEQ ID NO: 1470) (Fig. 3 A) and compared their effects on myogenic transdifferentiation (Fig. 3B-D). All threenovokines enhanced transdifferentiation efficiency. In contrast, neither natural ligands (FGF2, FGF7, FGF10, IL-6, EGF, BMP4, VEGF, TNF-a), designed FGFRl / 2c agonist C6-79C-mb7 nor HER2aff fused with other RTK minibinders (such as PDGFRmb, EGFRcmb, IGF2Rmb, TRKA Receptor mb and Insulin Receptor mb) increased transdifferentiation efficiency (Fig. 3C). These results suggest that HER2: FGFR heterodimerization activates a unique signaling pathway that facilitates myogenic fate conversion of fibroblasts (Fig. 3C, D). Direct muscle reprogramming was achieved with HER2 and FGFRl / 2c heterodimerization using two independent HER2 binders (HER2mb and HER2Affibody), despite their distinct binding surfaces and locations within the HER2 extracellular domain (Fig. 1C).

[0858] H2F treatment significantly enhanced myogenic conversion efficiency to about 56% compared with control (Fig. 3E). H2F-converted cells also exhibited two times greater multinucleation relative to control cells and formed significantly longer myotubes (~5-fold increase) (Fig. 3F, G). Additionally, myotubes treated with H2F or H2affF showed higher metabolic activity, characterized by a more elaborate mitochondrial network and increased Mito-Orange uptake. Bulk mRNA analysis showed increased expression of various muscle markers in H2F treated myotubes (Fig. 3H). Collectively, these findings indicate that H2F not only increases the efficiency of direct reprogramming but also promotes downstream myogenic differentiation. Myofiber maturation was significantly enhanced when the H2F treatment was extended from 4 days to 30 days, as evidenced by significantly increased formation of well-organized Z-bands of sarcomeric a-actinin in the transdifferentiated muscle cells (Fig. 31, J).

[0859] Collectively, these data indicate that H2F minibinders effectively promote the conversion of fibroblasts into myotubes and their subsequent maturation. Furthermore, these findings suggest that heterodimerization of HER2 and FGFR receptors activates a distinct pro-myogenic signaling program that is not induced by other HER2 receptor tyrosine kinase pairings or by stimulation of FGFR or HER2 alone.

[0860] HER2mb-FGFRmb induced HER2-FGFR pair drives downstream signaling

[0861] We performed quantitative imaging Forster Resonance Energy Transfer (QLFRET) experiments to assess the interaction between HER2 and FGFR1 in the presence of the synthetic ligand H2F

[0030] , Experiments were performed with truncated receptors, with the intracellular domains substituted with fluorescent proteins (Fig. 4A, B). HER2-mEYFP (donor) and FGFR-mCherry™ (acceptor) were co-expressed in individual CHO cells, andtheir expression levels were comparable in both H2F-treated and untreated conditions (Fig.

[0862] 4SB). In the presence of H2F, FRET efficiencies were elevated relative to the no-ligand control (Fig. 4A; in the latter case FRET efficiencies were still higher than baseline

[0031] , suggesting possible interactions between HER2 and FGFR1 in the absence of ligand.

[0863] Comparison of the deviation from “proximity FRET (Fig. 4A), shows a statistically-significant increase due to H2F, indicating that H2F stabilizes the HER2-FGFR heterocomplex.

[0864] To characterize the phosphoproteomic signatures induced by H2F, we treated CHO cells overexpressing human HER2 and FGFR1 with FGF, H2F, FBS or vehicle (no minibinder), and performed quantitative phosphopeptide enrichment analysis after LC-MS / MS. Across all conditions, 8,077 unique phosphosites were identified, of which 6,325 sites were sufficiently quantified and localized for further analysis (Fig-4SC). Comparative analysis revealed that while H2F and FGF shared a large subset of phosphorylated targets, including canonical MAPK pathway components such as MEK1 and ERK1 / 2, H2F also induced a unique phosphosignature (Fig. 4C, D). This suggests that H2F induces a partially overlapping but mechanistically distinct signaling program compared to native FGF ligands potentially reflecting altered receptor dimer topology and biased signal transduction downstream of HER2-FGFR1 heterodimers (Fig. 4D).

[0865] The dose-response curve for H2F-induced pERK revealed an EC so of 5.84 nM (Fig.

[0866] 4E), indicating high potency and specificity. To determine whether the kinase activities of HER2 and FGFR1 were required for H2F-dependent ERK activation, we examined CHO cells coexpressing kinase-null HER2 (HER2 K753M) with wild-type FGFR1, and kinase-null FGFR1 (FGFR1 K514M) with wild-type HER2. H2F failed to induce pERK, pAKT,pP38 in the presence of kinase-null FGFR1, whereas a robust pERK response persisted when HER2 was kinase-null (Fig. 4E, F). In CHO cells coexpressing wild-type HER2 and FGFR1, pretreatment with the HER2-specific kinase inhibitor Tucatinib15had no effect on H2F -induced pERK, whereas FGFR inhibitors PD 173074 markedly reduced ERK phosphorylation (Fig. 4G). These findings demonstrate that H2F-mediated pERK activation is strictly dependent on FGFR1 kinase activity but independent of HER2 kinase activity.

[0867] To further dissect whether proximity of both HER2 and FGFR receptors is needed to trigger downstream MAPK signaling, we examined three novokine constructs (H2F, FH2, and H2affF) designed to bind both HER2 and FGFRlc simultaneously. H2F novokines act as agonists, inducing robust phosphorylation of ERK, AKT, and p38 MAP kinases exclusively in cells co-expressing both HER2 and FGFR1, whereas cells expressing only HER2 or onlyFGFR1 showed no activation of these pathways (Fig. 4H, I). Furthermore, the signaling response was abolished by antagonistic HER2 or FGFR mini-binders (Fig. 4J). Similar results were obtained with two independently designed HER2 -binding constructs, strongly suggesting that H2F signaling is mediated by proximity -induced heterodimerization of HER2 and FGFRlc. In EA.hy926 cells (an immortalized human umbilical vein endothelial cell line), expressing native FGFR and HER2, treatment with the H2F, FH2mb, or H2affF constructs each induced phosphorylation of AKT (pAKT), whereas competition with either H2mb or Fmb markedly reduced pAKT levels (Fig. 4K). The signaling profile induced by the heterodimers H2F and FH2mb was distinct from that induced by the homodimers H2mb: H2mb (H2H2) and Fmb: Fmb (FF). Neither homodimer (H2H2 or FF) enhanced transdifferentiation efficiency or elicited pAKT signaling (Fig. 4J), in contrast to H2F. Taken together, these findings indicate that both receptors are required for the observed downstream signaling.

[0868] HER2-FGFR novokine represses PLCy activation and acts via MAPK arm of FGFR FGF and EGF activate a common set of intracellular signaling proteins (e.g. She, FRS2, PLCy, Grb2, Src, PI3K) upon ligand-induced tyrosine phosphorylation11,12. To investigate how H2F-induced HER2-FGFR1 heterodimers alter FGFR signaling, we examined FGFR1 phosphorylation after H2F treatment. H2F induced robust phosphorylation of FGFR1 at the activation loop tyrosines Y653 / Y654 - modifications that are essential for FGFR kinase activation- but did not induce phosphorylation of Y766. FGFR phosphorylation on Y653 / 654 is also abrogated by FGFRRTKs inhibitors (Fig.4E). the Phosphorylation of Y766 is required for PLCy recruitment and downstream Ca2+mobilization13,14. Consistent with this requirement, H2F stimulation failed to evoke any intracellular Ca2+transient (monitored by a fluorescent Ca2+indicator1), whereas FGF or a C6-79C-mb7(FGFRl / 2c designed agonist) agonist triggered a clear Ca2+response (Fig. 4L-N). Meanwhile, the maintained phosphorylation at Y653 / Y654 indicates preserved activation of the MAPK cascade.

[0869] Thus, H2F -induced HER2-FGFR1 heterodimers selectively propagate MAPK signaling while bypassing the PLCy / Ca2+ branch, effectively segregating the canonical FGFR outputs (Fig. 40). Collectively, these findings demonstrate a biased FGFR signaling response upon HER2-FGFR heterodimerization: FGFR kinase-dependent ERK / AKT pathways are engaged while the PLCy / Ca2+axis is excluded.H2F induced biased FGFR signaling supports pluripotency but not endothelial differentiation

[0870] The activation of FGFRl / 2c by H2F, which notably lacks the Ca2+signaling branch, enables us to pinpoint biological processes that rely on this specific FGFR modality. During vasculogenesis, both endothelial and perivascular cells arise from common mesodermal precursors, and FGF signaling plays a crucial role in their fate determination18,19. Our previous study demonstrated that selective activation of the FGFRl / 2c isoform using a designed FGF agonist (C6-79C-mb7) strongly promoted endothelial differentiation over perivascular fate, underscoring FGFRl / 2c’s role in endothelial lineage specification1. We therefore investigated whether the H2F novokine, which pairs FGFRl / 2c-splice variants with HER2 and activates only a partial FGFR pathway, could influence cell fate in a vascular differentiation model. To test this, we replaced FGF2 in the endothelial differentiation protocol with 100 nM of H2F, H2affF, or the monomeric FGFRl / 2c inhibitor mb7 (Fig. 5A). Furthermore, we showed that both FGFR and Her2 were expressed at this stage of differentiation. Flow cytometry analysis further showed that 75.48% of FGF2-treated cells expressed the endothelial marker VE-CAD, whereas with H2affF treatment only 21.14% expressed VE-CAD and with H2F treatment only 0.03% expressed VE-CAD (Fig. 5B).

[0871] Immunofluorescence imaging revealed that treatment with H2F and H2affF predominantly induced expression of the perivascular marker PDGFR-P (Fig. 5C). These findings suggest that novokines, by pairing FGFR with HER2, disrupt FGF-mediated endothelial differentiation and instead bias cell fate toward the perivascular lineage.

[0872] Since FGFR signaling is essential for human stem cell pluripotency20,21, we aimed to determine whether H2F-mediated biased FGFRl / 2c signaling, which selectively activates the MAPK pathway while bypassing the Ca2+ / PLCy axis, is sufficient to sustain pluripotency. First, to test if the FGFRl / 2c splice variant was sufficient for pluripotency, we treated hiPSCs with the FGFRl / 2c-specific antagonist mb7 (100 nM). This led to loss of Oct4 and downregulation of pluripotency markers, despite active b-isoforms, indicating FGFRl / 2c is essential for pluripotency. Conversely, replacing FGF with the FGFRl / 2c-specific agonist C6-79C-mb7 maintained 97-99% TRA-l-60+cells, outperforming FGF. These results demonstrate that FGFRl / 2c splice variant activation is both necessary and sufficient for maintaining human stem cell pluripotency. To further test H2F biased FGFRlc signaling in pluripotency, we treated WTC11 hiPSCs with C6-79C-mb7, mb7, FGF, or H2F in minimal media (E8) for 48 hours (Fig. 5D). Flow cytometry and immunostaining analyses showed that cells treated with C6-79C-mb7, FGF, or H2F maintained the expression of pluripotencytranscription factors (0CT4, Nanog) and the surface markers TRA1-60 / TRA1-81, whereas mb7-treated cells and FGF-deprived controls showed reduced marker expression (Fig. 5D, E). Seahorse analysis indicated that H2F-induced FGFR signaling did not alter mitochondrial respiration compared to the activation seen with FGF2 (Fig. 5F). The H2F treated iPSCs showed a positive correlation with FGF treated iPSCs for ectoderm and endoderm lineage differentiation (Fig-5G, H). These findings suggest that H2F-mediated signaling is sufficient to sustain hiPSC pluripotency, while FGFR dependent Ca2+ / PLCy signaling is dispensable (Fig. 51). FGF2 is a known component of stem cell maintenance media (e.g., mTeSR); our results indicate that specifically the FGFRlc / 2c-MAPK axis can uphold the pluripotency.

[0873] H2F enhances primary Myoblast fusion and maturation

[0874] In primary myoblasts from adult patients, H2F treatment significantly increased fusion efficiency and myotube coverage area compared to FGF treated controls, yielding thicker, more continuous fibers (Fig. 5J-N). Comparable effects were also observed in human iPSC-derived myoblasts. Unlike native FGF, which primarily drives myoblast proliferation, H2F elevated both fusion efficiency and a-actinin expression (Fig. 5M-N), underscoring a distinct mechanism of action. Sarcomere analysis further revealed that H2F promotes myotube maturation, with well-defined striations and physiological range for Z-disc spacing (median 1.87 pm) compared to the disorganized sarcomeres of the PBS-treated controls (median 1.28 pm) (Fig.5O-Q). Importantly, these results demonstrate thatH2F is not limited to fetal or iPSC-derived myogenic cells; it is equally effective in adult primary myoblasts, where regenerative potential is typically diminished. Thus, H2F emerges as a promising priming agent for autologous myoblast transplantation and a potential therapeutic to enhance muscle regeneration in settings of atrophy, injury, or aging (Fig. 5R).

[0875] Discussion

[0876] We find that pairing HER2 and the FGFRl / 2c using the novokine H2F drives fibroblast-to-muscle reprogramming. Similar results were obtained with H2F novokine-constructs carrying two distinct HER2 binding domains, strongly supporting that the observed activity reflects HER2 recruitment. FRET analysis confirmed that H2F mediates HER2-FGFR heterodimerization, a critical step in reprogramming. Phospho-proteomic profiling revealed that H2F engages signaling nodes such as various GAPs, GEFs, and MAPKs, overlapping partially with canonical FGF signaling. Mechanistic dissection showed that HER2-FGFR heterodimerization selectively bifurcates RTK signaling, activatingMAPK / AKT while bypassing PLCy. H2F relies on canonical FGFR Y653 / Y654 phosphorylation for downstream signaling. By activating the MAPK branch while avoiding Ca2+-dependent signaling, H2F enables dissection of FGFRl / 2c-dependent pathways across diverse biological context.

[0877] H2F-mediated FGFR activation is incompatible with endothelial differentiation, but crucial in myogenic differentiation, myofiber maturation, and pluripotency maintenance in human stem cells, suggesting that FGFR-dependent Ca2+signaling is dispensable for these processes. Single-cell RNA-seq from the Tabula Sapiens atlas identified human cell types co-expressing HER2 and FGFRl / 2c, pointing to potential in vivo contexts for H2F activity. As proof of concept, patient-derived primary myoblasts differentiated efficiently with H2F treatment, underscoring its promise for muscle regenerative medicine.

[0878] Our designed novokines now pioneer and establish the utility of the approach in natural cellular context. Novokines harness the ability to bring endogenously existing receptors together, creating unprecedented diversity of the receptor combinations and targeted cell types. With the rapidly expanding set of Al-designed binding domains, libraries of novokines can be systematically generated and screened across reprogramming and differentiation assays. As demonstrated with H2F, such synthetic ligands hold dual promise: they can serve as therapeutic leads for regenerative medicine and simultaneously uncover foundational principles of receptor wiring and signaling network logic.

[0879] References

[0880] (1) Edman, N. I.; Phal, A.; Redler, R. L.; Schlichthaerle, T.; Srivatsan, S. R.; Ehnes, D. D.; Etemadi, A.; An, S. J.; Favor, A.; Li, Z.; Praetorius, F.; Gordon, M.; Vincent, T.;

[0881] Marchiano, S.; Blakely, L.; Lin, C.; Yang, W.; Coventry, B.; Hicks, D. R.; Cao, L.; Bethel, N.; Heine, P.; Murray, A.; Gerben, S.; Carter, L.; Miranda, M.; Negahdari, B.; Lee, S.;

[0882] Trapnell, C.; Zheng, Y.; Murry, C. E.; Schweppe, D. K.; Freedman, B. S.; Stewart, L.; Ekiert, D. C.; Schlessinger, J.; Shendure, J.; Bhabha, G.; Ruohola-Baker, H.; Baker, D. Modulation of FGF Pathway Signaling and Vascular Differentiation Using Designed Oligomeric Assemblies. Cell 2024, 187 (14), 3726-3740.e43.

[0883] (2) Zhao, Y. T.; Fallas, J. A.; Saini, S.; Ueda, G.; Somasundaram, L.; Zhou, Z.; Xavier Raj, I.; Xu, C.; Carter, L.; Wrenn, S.; Mathieu, J.; Sellers, D. L.; Baker, D.; Ruohola-Baker, H. F-Domain Valency Determines Outcome of Signaling through the Angiopoietin Pathway. EMBO Rep 2021, 22 (12), e53471.(3) Cao, L.; Coventry, B.; Goreshnik, I.; Huang, B.; Sheffler, W.; Park, J. S.; Jude, K. M.; Markovic, I.; Kadam, R. U.; Verschueren, K. H. G.; Verstraete, K.; Walsh, S. T. R.; Bennett, N.; Phal, A.; Yang, A.; Kozodoy, L.; DeWitt, M.; Picton, L.; Miller, L.; Strauch, E - M.; DeBouver, N. D.; Pires, A.; Bera, A. K.; Halabiya, S.; Hammerson, B.; Yang, W.;

[0884] Bernard, S.; Stewart, L.; Wilson, I. A.; Ruohola-Baker, H.; Schlessinger, J.; Lee, S.;

[0885] Savvides, S. N.; Garcia, K. C.; Baker, D. Design of Protein-Binding Proteins from the Target Structure Alone. Nature 2022, 605 (7910), 551-560.

[0886] (4) Davis, R. L.; Weintraub, H.; Lassar, A. B. Expression of a Single Transfected cDNA Converts Fibroblasts to Myoblasts. Cell 1987, 51 (6), 987-1000.

[0887] (5) Tapscott, S. J.; Davis, R. L.; Thayer, M. J.; Cheng, P. F.; Weintraub, H.; Lassar, A. B. MyoDl: A Nuclear Phosphoprotein Requiring a Myc Homology Region to Convert Fibroblasts to Myoblasts. Science 1988, 242 (4877), 405-411.

[0888] (6) Kabadi, A. M.; Thakore, P. I.; Vockley, C. M.; Ousterout, D. G.; Gibson, T. M.; Guilak, F.; Reddy, T. E.; Gersbach, C. A. Enhanced MyoD-Induced Transdifferentiation to a Myogenic Lineage by Fusion to a Potent Transactivation Domain. ACS Synth Biol 2015, 4 (6), 689-699.

[0889] (7) Manandhar, D.; Song, L.; Kabadi, A.; Kwon, J. B.; Edsall, L. E.; Ehrlich, M.;

[0890] Tsumagari, K.; Gersbach, C. A.; Crawford, G. E.; Gordan, R. Incomplete MyoD-Induced Transdifferentiation Is Associated with Chromatin Remodeling Deficiencies. Nucleic Acids Res 2017, 45 (20), 11684-11699.

[0891] (8) Abdel-Raouf, K. M. A.; Rezgui, R.; Stefanini, C.; Teo, J. C. M.; Christoforou, N. Transdifferentiation of Human Fibroblasts into Skeletal Muscle Cells: Optimization and Assembly into Engineered Tissue Constructs through Biological Ligands. Biology (Basel) 2021, 10 (6), 539.

[0892] (9) Lemmon, M. A.; Bu, Z.; Ladbury, J. E.; Zhou, M.; Pinchasi, D.; Lax, I.; Engelman, D. M.; Schlessinger, J. Two EGF Molecules Contribute Additively to Stabilization of the EGFR Dimer. EMBO J 1997, 16 (2), 281-294.

[0893] (10) Bennett, N. R.; Coventry, B.; Goreshnik, I.; Huang, B.; Allen, A.; Vafeados, D.; Peng, Y. P.; Dauparas, J.; Baek, M.; Stewart, L.; DiMaio, F.; De Munck, S.; Savvides, S. N.; Baker, D. Improving de Novo Protein Binder Design with Deep Learning. Nat Commun 2023, 74 (1), 2625.

[0894] (11) Ferguson, H. R.; Smith, M. P.; Francavilla, C. Fibroblast Growth Factor Receptors (FGFRs) and Noncanonical Partners in Cancer Signaling. Cells 2021, 10 (5), 1201.(12) Boonstra, J.; Rijken, P.; Humbel, B.; Cremers, F.; Verkleij, A.; van Bergen en Henegouwen, P. The Epidermal Growth Factor. Cell Biol Int 1995, 19 (5), 413-430.

[0895] (13) Farrell, B.; Breeze, A. L. Structure, Activation and Dysregulation of Fibroblast Growth Factor Receptor Kinases: Perspectives for Clinical Targeting. Biochem Soc Trans 2018, 46(6), 1753-1770.

[0896] (14) Cross, M. J.; Hodgkin, M. N.; Roberts, S.; Landgren, E.; Wakelam, M. J.; Claesson-Welsh, L. Tyrosine 766 in the Fibroblast Growth Factor Receptor-1 Is Required for FGF-Stimulation of Phospholipase C, Phospholipase D, Phospholipase A(2), Phosphoinositide 3-Kinase and Cytoskeletal Reorganisation in Porcine Aortic Endothelial Cells. J Cell Sci 2000, 113 (Pt 4), 643-651.

[0897] (15) Olson, D.; Taylor, J.; Willis, K.; Hensley, K.; Allred, S.; Zaval, M.; Farr, L.; Thurman, R.; Jain, N.; Hein, R.; Ulrich, M.; Peterson, S.; Kulukian, A. HER2-Selective and Reversible Tyrosine Kinase Inhibitor Tucatinib Potentiates the Activity of T-DM1 in Preclinical Models of HER2 -Positive Breast Cancer. Cancer Res Commun 2023, 3 (9), 1927- 1939.

[0898] (16) Mohammadi, M.; Froum, S.; Hamby, J. M.; Schroeder, M. C.; Panek, R. L.; Lu, G. H.; Eliseenkova, A. V.; Green, D.; Schlessinger, J.; Hubbard, S. R. Crystal Structure of an Angiogenesis Inhibitor Bound to the FGF Receptor Tyrosine Kinase Domain. EMBO J 1998, 77(20), 5896-5904.

[0899] (17) Mohammadi, M.; McMahon, G.; Sun, L.; Tang, C.; Hirth, P.; Yeh, B. K.; Hubbard, S. R.; Schlessinger, J. Structures of the Tyrosine Kinase Domain of Fibroblast Growth Factor Receptor in Complex with Inhibitors. Science 1997, 276 (5314), 955-960.

[0900] (18) Javerzat, S.; Auguste, P.; Bikfalvi, A. The Role of Fibroblast Growth Factors in Vascular Development. Trends Mol Med 2002, 8 (10), 483-489.

[0901] (19) Vodyanik, M. A.; Yu, J.; Zhang, X.; Tian, S.; Stewart, R.; Thomson, J. A.; Slukvin, 1. 1. A Mesoderm-Derived Precursor for Mesenchymal Stem and Endothelial Cells. Cell Stem Cellis, 7(6), 718-729.

[0902] (20) Lanner, F.; Rossant, J. The Role of FGF / Erk Signaling in Pluripotent Cells. Development 2010, 737 (20), 3351-3360.

[0903] (21) Ware, C. B.; Nelson, A. M.; Mecham, B.; Hesson, J.; Zhou, W.; Jonlin, E. C.; Jimenez-Caliani, A. J.; Deng, X.; Cavanaugh, C.; Cook, S.; Tesar, P. J.; Okada, J.;

[0904] Margaretha, L.; Sperber, H.; Choi, M.; Blau, C. A.; Treuting, P. M.; Hawkins, R. D.; Cirulli, V.; Ruohola-Baker, H. Derivation of Naive Human Embryonic Stem Cells. Proc Natl Acad Sci USA 2014, 777 (12), 4484-4489.(22) Ben-Sasson, A. J.; Watson, J. L.; Sheffler, W.; Johnson, M. C.; Bittleston, A.; Somasundaram, L.; Decarreau, J.; Jiao, F.; Chen, J.; Mela, I.; Drabek, A. A.; Jarrett, S. M.; Blacklow, S. C.; Kaminski, C. F.; Hura, G. L.; De Yoreo, J. J.; Kollman, J. M.; Ruohola-Baker, H.; Delivery, E.; Baker, D. Design of Biologically Active Binary Protein 2D Materials. Nature 2021, 589 (7842), 468-473.

[0905] (23) Divine, R.; Dang, H. V.; Ueda, G.; Fallas, J. A.; Vulovic, I.; Sheffler, W.; Saini, S.; Zhao, Y. T.; Raj, I. X.; Morawski, P. A.; Jennewein, M. F.; Homad, L. J.; Wan, Y -H.; Tooley, M. R.; Seeger, F.; Etemadi, A.; Fahning, M. L.; Lazarovits, J.; Roederer, A.; Walls, A. C.; Stewart, L.; Mazloomi, M.; King, N. P.; Campbell, D. J.; McGuire, A. T.; Stamatatos, L.; Ruohola-Baker, H.; Mathieu, J.; Veesler, D.; Baker, D. Designed Proteins Assemble Antibodies into Modular Nanocages. Science 2021, 372 (6537), eabd9994.

[0906] (24) Lutz, I. D.; Wang, S.; Norn, C.; Courbet, A.; Borst, A. J.; Zhao, Y. T.; Dosey, A.; Cao, L.; Xu, J.; Leaf, E. M.; Treichel, C.; Litvicov, P.; Li, Z.; Goodson, A. D.; Rivera-Sanchez, P.; Bratovianu, A.-M.; Baek, M.; King, N. P.; Ruohola-Baker, H.; Baker, D. Top-down Design of Protein Architectures with Reinforcement Learning. Science 2023, 380 (6642), 266-273.

[0907] (25) Albanell, J.; Baselga, J. Trastuzumab, a Humanized Anti-HER2 Monoclonal Antibody, for the Treatment of Breast Cancer. Drugs Today (Bare) 1999, 35 (12), 931-946.

[0908] (26) Krzyscik, M. A.; Por^bska, N.; Opalinski, L.; Otlewski, J. Targeting HER2 and FGFR-Positive Cancer Cells with a Bispecific Cytotoxic Conjugate Combining Anti-HER2 Affibody and FGF2. Int J Biol Macromol 2024, 254 (Pt 1), 127657.

[0909] (27) Smith AS, Luttrell SM, Dupont JB, Gray K, Lih D, Fleming JW, Cunningham NJ, Jepson S, Hesson J, Mathieu J, Maves L, Berry BJ, Fisher EC, Sniadecki NJ, Geisse NA, Mack DL. High-throughput, real-time monitoring of engineered skeletal muscle function using magnetic sensing. J Tissue Eng. 2022 Sep 2;13:20417314221122127.

[0910] (28) Wang G, Luo X, Gu R, Yang S, Qu Y, Zhai S, Zhao Q, Li K, Zhang S. PyMIC: A deep learning toolkit for annotation-efficient medical image segmentation. Comput Methods Programs Biomed. 2023 Apr;231: 107398.

[0911] (29) Haghofer A, Fuchs-Baumgartinger A, Lipnik K, Klopfleisch R, Aubreville M, Scharinger J, Weissenbock H, Winkler SM, Bertram CA. Histological classification of canine and feline lymphoma using a modular approach based on deep learning and advanced image processing. Sci Rep. 2023 Nov 9; 13(1): 19436.(30) Chen L, Novicky L, Merzlyakov M, Hristov T, Hristova K. Measuring the energetics of membrane protein dimerization in mammalian membranes. J Am Chem Soc. 2010 Mar 17; 132(10):3628-35. doi: 10.1021 / ja910692u. PMID: 20158179

[0912] (31) King C, Sarabipour S, Byrne P, Leahy DJ, Hristova K. The FRET signatures of noninteracting proteins in membranes: simulations and experiments. Biophys J. 2014 Mar 18; 106(6): 1309-17. doi: 10.1016 / j.bpj.2014.01.039. PMID: 24655506

[0913] (32) Zhao Y, Ogishi M, Pal A, Su LL, Tao P, Jiang H, Rodriguez GE, Chen X, Sun Q, Rysavy LW, Limsuwannarot S, Waghray D, Kalbasi A, Garcia KC. Expanding the cytokine receptor alphabet reprograms T cells into diverse states. Nature. 2025 Aug 13. doi:

[0914] 10.1038 / s41586-025-09393-l. Epub ahead of print. PMID: 40804519.

[0915] (33) Chen G, Gulbranson DR, Hou Z, Bolin JM, Ruotti V, Probasco MD, Smuga-Otto K, Howden SE, Diol NR, Propson NE, Wagner R, Lee GO, Antosiewicz -Bourget J, Teng JM, Thomson JA. Chemically defined conditions for human iPSC derivation and culture. Nat Methods. 2011 May;8(5):424-9. doi: 10.1038 / nmeth.l593. Epub 2011 Apr 10. PMID:

[0916] 21478862; PMCID: PMC3084903.

[0917] (34) Hughes CS, Moggridge S, Muller T, Sorensen PH, Morin GB, Krijgsveld J. Single¬ pot, solid-phase-enhanced sample preparation for proteomics experiments. NatProtoc. 2019 Jan;14(l):68-85. doi: 10.1038 / s41596-018-0082-x. PMID: 30464214.

[0918] (35) Rappsilber J, Mann M, Ishihama Y. Protocol for micro-purification, enrichment, pre¬ fractionation and storage of peptides for proteomics using StageTips. Nat Protoc.

[0919] 2007;2(8): 1896-906. doi: 10.1038 / nprot.2007.261. PMID: 17703201.

[0920] (36) Benjamini, Yoav, and Yosef Hochberg. “Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing.” Journal of the Royal Statistical Society. Series B (Methodological), vol. 57, no. 1, 1995, pp. 289-300. JSTOR,.

[0921] (37) King C, Raicu V, Hristova K. Understanding the FRET Signatures of Interacting Membrane Proteins. J Biol Chem. 2017 Mar 31;292(13): 5291 -5310. doi:

[0922] 10.1074 / jbc. Ml 16.764282. Epub 2017 Feb 9. PMID: 28188294; PMCID: PMC5392676.

[0923] Materials and Methods

[0924] Cell culture- CHO cells, immortalized HUVEC cells, HFF-MyoD, Primary human myoblasts IPSCs

[0925] HFF-MyoD cells- HFF-MyoD cells were maintained in growth media (DMEM with 4.5g / L glucose and Glutamax™ [Gibco #10566024], 10% fetal bovine serum (FBS) [BioWest #S 1620], 1% penicillin / streptomycin (P / S) [Gibco #15140122], 1% NEAA [Gibco#11140050]) at 37°C and 5% CO2. At 80% confluency HFF-MyoD cells were passaged and seeded at a 1:10 ratio. Chinese hamster ovary (CHO) cells - CHO cells were maintained in CHO growth media (F12K [ATCC #30-2004; Gibco #21127022], 10% FBS, 1% P / S) at 37°C and 5% CO2. At 90% confluency CHO cells were passaged and seeded at a 1:10 ratio.

[0926] Continued background selection of CHO overexpression (OE) lines was maintained through addition of 5 pg / mL blasticidin [Company, catalog no] or 10 pg / ml puromycin [Gibco #A1113803] to the CHO growth media depending on the plasmid used. Immortalized human umbilical vein endothelial cells (HUVECs) - EA.hy926 cells were maintained in growth media (DMEM with 4.5g / L glucose and Glutamax™, 10% FBS, 1% P / S, 1% NEAA) at 37°C and 5% CO2. At 80% confluency EA.hy926 cells were passaged and seeded at a 1:10 ratio. Primary human skeletal muscle myoblasts [Cook Myosite #SK-1111] and IPSC derived myoblasts

[0027] were maintained in Skeletal muscle growth media (SKGM) consisting of 10% FBS [Hycone #SH30071.03), lOOnM Insulin [Lonza #BE02-033E20], 40ng / mL FGF-2[R& D Systems #3718-FB] & IpM Dexamethonsone [Sigma #D4902] in DMEM with 4.5g / L glucose and Glutamax™ [Gibco #10566024],

[0927] Generation of Skeletal Muscle Myoblasts from iPSCs

[0928] iPSC-derived myoblasts (iPSC-MBs) were generated from the UC3-4 iPSC line using a multi-stage protocol adapted from previous work

[0027] , iPSCs were plated at 15,000 cells / cm2in mTeSR Plus™ (Stem Cell Tech) and grown to -40% confluency in 6-well plates. Myogenic induction was initiated (day 0) with differentiation medium containing DMEM / F12 (Gibco #10565018), lx non-essential amino acids (Gibco #11140050), lx Insulin-Transferrin-Selenium (Gibco #41400045), 3 pM CHIR99021 (Axon #1386), and 0.2 pg / mL LDN193189 (Miltenyi Biotec #130-106-540). On day 2, medium was supplemented with 20 ng / mL bFGF (R& D Systems #3718-FB). On day 5, CHIR99021 and ITS were removed and 15% KSR (Gibco #10828028), 2 ng / mL IGF-1 (R& D Systems #291-G1), and 10 ng / mL HGF (R& D Systems #294-HGN) were added. On day 7, bFGF and LDN193189 were removed. Medium was changed every other day until day 28. At day 28 post-induction, a single confluent cell sheet was mechanically dissociated into small clumps and replated onto a Matrigel™-coated T225 flask for expansion in SKGM, with feeding every 2-3 days. At day 32, cells were lifted, filtered through a 40pm filter and subjected to fluorescence-activated cell sorting (FACS) to purify for positive Nerve derived growth factor (NGFR+) population [Biolegend #345108], All downstream experiments used cells between passages 4 and 5.Screening novokines in Myogenic transdifferentiation assay

[0929] In a 96 well plate, 10A3 per well HFF-tet-on inducible MyoD cells in DMEM with 10% FBS were seeded. Post 2 days of seeding, for the next 4 days cells were treated with 2 nM doxycycline [company, catalog number] in SMM media. The following 4 days cells are incubated with lOOnM of each novokine. Cells were fixed in 4% paraformaldehyde [Electron Microscopy Sciences #15710, diluted in phosphate buffered saline (PBS)] for 15 min, washed with PBS, and stained for Desmin / MHC, followed by imaging at Leica spinning disk microscope, stitched (4x4) images of each 96 well at 10X was captured. In Fiji ImageJ, the desmin intensity area was thresholded and the intensity was measured as the desmin positive area.

[0930] Screening myogenic fusion of Primary and IPSC derived myoblasts

[0931] Primary and iPSC-derived myoblasts were seeded In 24 well plate polymer bottom plates [Cellvis #P24-1.5P] at 15,000 / cm2and grown in SKGM until confluent. To initiate myogenic differentiation (day 0), media was changed to DMEM high glucose supplement with 2% Horse serum [Gibco #16050114] + / - H2F of varying concentrations. The media was changed every 48 hours. On Day 6, cultures were fixed in 4% paraformaldehyde for 20 minutes at room temperature. Cells were then permeabilized and blocked for 1 hour using a blocking buffer consisting of 0.2% Triton X-100, 1% BSA, and 5% Donkey Serum in PBS. Following blocking, cells were incubated overnight at 4°C with a primary antibody against sarcomeric a-actinin (R& D Systems, #MAB9830, at 1:500 dilution. After washing, cells were incubated with a fluorescently labeled secondary antibody and counterstained with DAPI to visualize nuclei. Images were acquired on a Yokogawa W 1 spinning disk confocal microscope at 20x magnification. For each well, a large 4x4 image panel was captured to provide a comprehensive view of the culture. An in-house ImageJ macro was used for automated, unbiased quantification. First, the LABKIT™ plugin

[0028] was used to segment a myotube mask from the a-actinin channel. Next, the StarDist plugin

[0029] was used to label the nuclei from the DAPI channel. A nucleus was classified as "fused" if it met two criteria: (1) area > 40 pm2, and (2) >90% of its area overlapped with the myotube mask. Fusion efficiency was calculated as the percentage of fused nuclei relative to the total nuclei per image.

[0932] Design of HER2 minibinderThe top 96 hits from sequencing were expressed and purified from 1 mL E. coli cultures in 96 well plate format, followed by screening on the Octet™ / BLI system using a two-point titration (1000 nM and 200 nM). The top 10 binders were selected based on binder saturation at both concentrations as well as the flatness of dissociation traces observed. For these 10 binders, we performed a three-fold titration series from 1000 nM to 4 nM, which demonstrated low picomolar affinities via global kinetic fitting (Supplementary FigurelB). However, due to the small size (6 kDa) of the binders, the signals were relatively small when binding to the much larger, glycosylated HER2 target protein (70-90 kDa). To address this, the top 10 binders were expressed as GFP fusion proteins (~33 kDa), purified from 50 mL E. coli cultures, and reassessed using Octet™ / BLI to enhance the signal-to-noise ratio. The GFP fusion proteins produced significantly larger signals, consistent with the original kinetic fitting results, confirming low pM affinities with near-flat dissociation rates over the duration of the experiment (FigurelC). The top binder exhibited a single, monodisperse peak in size exclusion chromatography (SEC), both with and without the GFP fusion tag (data not shown). Structural analysis of the design model revealed key interactions with HER2, where PHE51 and TYR32 from the minibinder fit into complementary hydrophobic pockets of the target (FigurelC).

[0933] Ca2+assay

[0934] CHO-hHER2-hFGFRlc cells were seeded on 96-well flat bottom microplates [Corning #3603] and grew to 70-80% confluence. Cells were starved in serum-free Fl 2-K medium for 3 hours. Following starvation, the cells were incubated in serum-free media containing 5mM Calbryte™ 520 AM fluorescent intracellular calcium indicator [AAT Bioquest, #20651] for 30 min at 37°C. Cells were washed 3X with serum-free media and treated with various concentrations of recombinant FGF2 (with or without 40mg / mL Heparin [Iduron, #H010]) or designed scaffolds. Confocal live imaging was done on a Nikon Yokogawa W1 spinning disk confocal microscope using a 20X objective. Parameters for each live frame: Excitation / Emission filters for GFP fluorescence, Exposure time of 150ms, Acquisition rate of 5 sec / frame, and total recording time of 20 minutes (5 min baseline recording + 15 min ligand treatment time). Images were processed with Fiji software distribution of ImageJ vl.52i and frame-by-frame cellular fluorescence intensity was tracked and quantified with CellProfiler™. Dose-specific average calcium release was calculated by tracking each individual cell’s response during the recording time and computing the meanpeak fluorescence achieved by all cells in the frame. An average of 50-100 cells were tracked per recording.

[0935] In vitro differentiation of endothelial cells

[0936] Briefly, hiPSCs (WTC-11 human induced pluripotent stem cells) [Coriell, #GM25256] were seeded on 24-well plates coated with growth factor-reduced Matrigel™ [Corning, #356231] and cultured in mTeSRl stem cell medium [StemCell Technologies, #85850] until cells reach confluence with media changes daily. One day before differentiation (deemed Day (-1)), cells were pre-treated with mTeSRl supplemented with IpM of GSK3 -Inhibitor (CHIR99021) [Cayman Chemicals, #13122], On the first day of differentiation (DO), stem cell media was replaced with cardiogenic mesoderm media consisting of RPMI 1640 Medium [Thermo, #11875093] supplemented with 1XB27(-) [Fisher Scientific, #A1895601], 100 ng / mL Activin A [PeproTech, #120-14P] and IX Matrigel™ for 17 hrs. The next day, media was replaced with RPMI supplemented with IpM of GSK3 -Inhibitor (CHIR99021), B27 (-), and 5 ng / mL bone morphogenetic protein-4 (BMP-4) [R& D systems, #314-BP-010] for 24 hours. On Day 2 of differentiation, cells were washed with IX PBS and media was replaced with vascular differentiation media consisting of StemPro™ [Thermo Fisher, #10639011] supplemented with IX Glutamax™, IX Penicillin-Streptomycin, 300 ng / mL vascular endothelial growth factor (VEGF) [R& D systems, #293-VE-050], 10 ng / mL BMP -4, 5 ng / mL FGF2, 50 ug / mL Ascorbic Acid [Sigma-Aldrich, #A8960], and 40 pM monothioglycerol (MTG) [Sigma-Aldrich, #M6145], On Day 5, cells were dissociated with Accutase™ [Thermo, #A1110501] and replated on 12-well 0.1% gelatin-coated tissue culture dishes in endothelial growth media (EGM) consisting of EGM basal media [Lonza, #CC-3121] supplemented with 20 ng / mL VEGF, 20 ng / mL FGF2 and IpM GSK3 -Inhibitor (CHIR99021). EGM media was replaced every 48 hours until the final harvest at Day 14.

[0937] Flow cytometry

[0938] Cells derived using FGF2, H2F, and mb7 (at Day 14 of differentiation) were harvested as a single cell suspension, adjusted to a concentration of 0.5 x 106cells / mL in ice-cold FACS Buffer (IX PBS, 1% BSA, 0.1% sodium azide [Millipore- Sigma, #26628-22-8]), and blocked for 30 minutes on ice. Cells were washed 3 times in IX PBS by centrifugation at 1500 rpm for 5 minutes each, following which primary (labeled) antibodies were added at a dilution of 1: 100 in FACS buffer for 1 hr at 4C in the dark: VE-Cadherin-APC [eBioscience,#17-1449-42], PDGFR-B-APC [BioLegend, #323608], Cells were washed 3 times in IX PBS by centrifugation at 1500 rpm for 5 minutes each and resuspended in lOOuL of ice-cold FACS Buffer for subsequent analysis. Unstained controls were included. Samples were run on a FACSCanto™ II flow cytometer (BD Biosciences) and recorded events were analyzed using the flowCore™ package for R. FSC and SSC (Unstained control) were used for size gating. Events were analyzed as the percentage of cells positive for the given panel of markers.

[0939] Immunostaining of differentiated iPSCs

[0940] For immunofluorescence imaging of differentiated iPSCs, cells were seeded on glass coverslips coated with 0.1% gelatin on Day 5 and cultured until confluency on Day 14 following the process described above. The cells were then fixed in 4% paraformaldehyde (PF A) for 15 minutes. The fixed cells were washed three times for 5 min each in IX PBS before blocking for 1 hr with 3% BSA and 0.1% Triton X-100 in IX PBS while on nutation. Primary antibody incubation was carried out at a 1: 100 dilution in blocking buffer overnight at 4C: CD31 (Cell Signaling, Catalog #3528), and PDGFR-B (Cell Signaling, Catalog #3169). Following overnight incubation, the cells were washed three times for 5 min each in IX PBS while on nutation. The cells were then incubated with secondary antibodies (Invitrogen, A21050 and Invitrogen, Al 1008; 1:200 each) diluted in blocking buffer for 1.5 hrs at 37°C. Secondary antibodies were then removed, and cells were washed three times for 10 min each in IX PBS on nutation. Coverslips were sealed using VECTASHIELD™ with DAPI [Vector laboratories, #H-2000-2] upside-down on glass slides for analysis in confocal microscopy. Images were taken on a Nikon Yokogawa™ W1 spinning disk confocal microscope using a 20X objective.

[0941] Pluripotency assays

[0942] hiPSCs (WTC-11 human induced pluripotent stem cells) [Coriell, #GM25256] were seeded onto 12-well plates pre-coated with growth factor-reduced Matrigel™ [Coming, #356231] at a density of 20,000 cells per well, in mTeSRl medium supplemented with 10 pM Rock inhibitor (Y-27632, Tocris Bioscience). The following day, the media was replaced with E8 basal media consisting of DMEM / F12 supplemented with L-ascorbic acid-2-phosphate magnesium (64 mg / L), sodium selenite (14 pg / L), insulin (19.4 mg / L), transferrin (10.7 mg / L), FGF2 (100 pg / L), and TGF-pi (2 pg / L)

[0033] , For experimental conditions, FGF2 was replaced with 100 nM of the FGFRlc-specific inhibitor minibinder (mb7) or theFGFR-HER2 novokine (H2F). Cells were cultured in these experimental conditions for 48 hours before harvesting for analysis.

[0943] FRET imaging and data analysis

[0944] FRET imaging was then performed using a Leica SP8 confocal microscope as discussed in detail previously. The donor scan excited YFP at 488 nm and its emission was collected between 500 and 540 nm. The acceptor scan excited the acceptor at 552 nm, and its emission was collected from 590 to 700 nm. For the FRET scan, YFP was excited, and mCherry™ emission was detected in the 590 to 700 nm range. Cells were imaged in the absence and in the presence of 500 nM of H2F protein. One or two small (~5 mm) regions of the plasma membrane were selected per cell for FRET analysis. The FRET efficiency, the donor concentration (HER- YFP), and acceptor concentration (FGFRlc-mCherry™) were measured in each region. For both conditions, with and without H2F protein, the FRET efficiency as a function of the acceptor concentration, the donor concentration as a function of the acceptor concentration, and the deviation from the proximity for each point were plotted using the software package Origin. The deviation from proximity FRET (Fig. 4A) was performed using an unpaired t-test with the Prism software.

[0945] Phosphoproteomics Analysis

[0946] CHO-hHER2-hFGFRlc cells were seeded on 6-well plates and grown to 80% confluence and serum starved for four hours prior to treatment. Cells were treated for 15 minutes with either lOOnM of the FGFR-HER2 novokine, 1 nM of FGF, media containing 10% fetal bovine serum, or serum -free media with PBS. After treatment, cells were washed twice with PBS and lysis buffer (8M urea, 50mM NaCl, 200 mM EPPS [pH 8.5], Roche phosSTOP™ phosphatase inhibitor tablets, and Roche protease inhibitor tablets) was added lyse cells prior to processing. For phosphoproteomics, 100 pg of protein lysate for each sample was processed. Lysates were reduced (5 mM TCEP [Sigma Aldrich], 20 min) and alkylated (10 mM iodoacetamide [Sigma Aldrich]); excess iodoacetamide was quenched with the addition of 10 mM dithiothreitol [Sigma Aldrich], Proteins were extracted using the SP3 protocol

[0034] prior to resuspension in 200 mM EPPS (pH 8.5) and digestion with LysC Wako) for 18 hours with vortexing followed by trypsin digestion for 6 hours at 37°C at 200 rpm. Peptides were labeled with TMTpro reagents [Thermo Fisher Scientific] for 1 hour at room temperature on the SP3 beads, then quenched with 5% hydroxylamine and incubation for 15 minutes at room temperature. TMTpro-labeled peptides were cleaned using C18SepPaks™ (Waters) and the resulting eluate was dried to completion in a speedvac.

[0947] Phosphopeptide enrichment was performed using Fe-NTA magnetic beads (PureCube™ Fe-NTA MagBeads). Phosphopeptides were cleaned using Cl 8 STAGE-tips

[0035] and eluates were dried to completion and frozen at -80°C prior to analysis by LC-MS / MS.

[0948] Phosphopeptides were resuspended in 5% acetonitrile and 2% formic acid then analyzed using an SPS-MS3 method on an Orbitrap Tribrid Eclipse™ and a 180 minute method with a flow rate of 300 nl / min on a C18 analytical column (15 cm Odyssey, ionOpticks). A gradient was applied starting in 100% solvent A (0.125% formic acid), then increasing from 4% solvent B (95% acetonitrile, 0.125% formic acid) to 20% solvent B after 165 minutes. MSI spectra (Orbitrap Resolution = 120,000; maximum injection time = 50 ms; normalized AGC = 200%; RF lens % = 30) were collected for precursor detection cycling between three FAIMS CVs (-40, -60, -80) [REF], MS2 spectra were collected for precursors between 300-1500 m / z using quadrupole isolation, dynamic exclusion (time delay = 90 seconds, window = 10 ppm) and intensity filtering for ion trap spectra (>5000). Ion trap MS2 spectra (CID energy % = 35; multistage activation = true; neutral loss mass = 97.9763; maximum injection time = 35 ms; normalized AGC % = 250) were used for peptide identification. Orbitrap MS3 spectra (HCD energy % = 45; first m / z = 110; Orbitrap Resolution = 50,000; maximum injection time = 86 ms; normalized AGC % = 250) were used for TMTpro reporter ion quantification. Phosphopeptides were identified using the Comet search algorithm and filtered to a 1% protein and peptide false discovery rate using linear discriminant analysis and the rules of protein parsimony.

[0949] Statistical analysis- Two-sided Student’s t-test was performed to determine p-value. For phosphoproteomics analyses, Welch’s t-tests were used to assess statistical significance followed by Benjamini -Hochberg procedure

[0036] to correct for multiple hypothesis testing.

[0950] Example 2

[0951] Abstract

[0952] Direct somatic cell reprogramming holds promise for regenerative medicine but remains limited by inefficient conversion and incomplete functional maturation. Here we screen a library of Al-designed receptor-modulatory minibinders and identify a synthetic protein cocktail, C6DPC, that robustly drives human fibroblast-to-muscle transdifferentiation. In an inducible MyoD system, C6DPC markedly increases conversion efficiency and promotes myotube fusion, structural maturation, and metabolic remodeling with elevatedmitochondrial respiration. Mechanistically, C6DPC rewires extracellular signaling by activating pro-myogenic FGFRl / 2c pathways while suppressing inhibitory ALK1 and TGFBR2 signaling that otherwise divert cells toward fibrotic states. Targeted depletion of ALK1 via minibinder-mediated lysosomal degradation is sufficient to lower the reprogramming barrier and enhance myogenic conversion. Importantly, C6DPC-generated muscles display superior functional maturation, producing greater twitch and tetanic forces in engineered tissues derived from both wild-type and dystrophin-deficient (DMD) human cells. These findings show that programmable synthetic ligands can control the signaling logic of transdifferentiation and generate functionally stronger muscle, establishing a strategy for engineering cell fate transitions in regenerative medicine.

[0953] Introduction

[0954] Skeletal muscle homeostasis is naturally maintained by satellite cells, but their regenerative capacity declines precipitously with age or due to irreversible trauma, surgery, and genetic disorders such as Duchenne Muscular Dystrophy (DMD). While dedifferentiation into induced myogenic progenitor cells (iMPCs) using factors like FRC (Forskolin, CHIR99021, RepSOX) has been explored, these ceils remain highly proliferative and carry risks associated with multi-stage transitions (Yagi et al., 2021; Kim et al., 2022).

[0955] Consequently, optimizing direct reprogramming remains the most desirable strategy for rapid, safe muscle repair.

[0956] Direct cellular reprogramming represents a transformative frontier for regenerative medicine, offering a path to bypass the complexities of pluripotency while providing a source of patient-specific functional cells. The conversion of fibroblasts into myogenic lineages via the ectopic expression of MyoD was the seminal demonstration of cell fate plasticity (Weintraub et al., 1987; (Weintraub et al., 1987; Morris & Daley, 2013). However, despite decades of research, the therapeutic application of direct reprogramming is still hampered by low conversion efficiency (Cacchiarelli et al., 2015; Kim et al., 2022). One major barrier is restricted chromatin accessibility, which limits MyoD binding to key regulatory elements and results in incomplete transcriptional activation of the myogenic program (Manandhar et al., 2017; Kolundzic et al., 2018; Liu et al., 2018). In addition, extracellular signaling pathways such as BMP, Wnt, and insulin signaling influence the efficiency and stability of myogenic conversion (Rochat et al., 2004; Kodaka et al., 2017; Shirakawa et al., 2022).

[0957] Recent breakthroughs in Al-based molecular modeling, such as AlphaFold™ and RFDiffusion™, have introduced a new class of therapeutic agents: synthetic minibinders.These de novo designed proteins can be engineered with exquisite specificity down to the receptor isoform level and can function as either potent antagonists in monomeric form or "superagonists" when organized into high-valency oligomeric scaffolds. Unlike natural ligands, these minibinders can be displayed on rigid or flexible scaffolds to precisely tune signaling geometries.

[0958] In this study, we leverage these Al-designed proteins to overcome the efficiency bottlenecks of myogenic conversion. We report that a strategic combination of minibinders specifically targeting the activation of FGFRlc alongside the inhibition of TGFBRII and ALK1R, significantly ameliorates the efficiency and maturation of human fibroblast-derived muscle cells. Our results demonstrate that these synthetic protein cocktails not only refine the reprogramming process but also facilitate the functional rescue of DMD phenotypes, marking a new era in precision regenerative therapy.

[0959] Results

[0960] MyoD mediated direct reprogramming of fibroblast

[0961] In this study we are utilizing designed proteins to target the RTKs / RS / TKs to make the human myogenic cell fate conversion more efficient. We chose a neonatal skin fibroblast HFF (human Foreskin Fibroblast) to generate a HFF-inducible MyoD (HFFiMYOD) line under regulation of tet-on promoter (Fig. 6A). We found the expression of nuclear MyoD upon doxycycline treatment for 24 hours. However, a continuous overexpression MyoD in HFFiMYOD for d7 and dl4 did not show complete conversion of fibroblast to myogenic lineage. The transformed cells that failed to express muscle marker (cluster2) clustered separately from the resting fibroblast (cluster 1) suggesting negligible non-recombinant HFF in the HFFiMYOD line. The cluster 2 also does not express the myobfibroblast markers and cluster 3 is the cluster positive with muscle markers (Fig. 6B). The transcriptomic analysis reveals three distinct cell populations emerging from the transdifferentiation protocol. We identified a population of resting fibroblasts (cluster 1) maintaining baseline markers (MME, CXCL12) and a separate population of activated myofibroblasts (cluster 2) enriched for fibrotic markers (FN1, IGFBP5) (Fig. 6C). Most importantly, the emergence of Cluster 3 confirms successful reprogramming, as evidenced by the robust expression of definitive muscle structural and functional genes (ACTC1, CHRNA1), suggesting that while a subset of cells successfully transdifferentiates, another subset diverts towards an activated myofibroblast state (Fig. 6C).We analyzed a set of barrier genes (ID1, ID2, ID3, TGFB1, TGFB2, TGFBR1, TWIST1, TWIST2, ZEB1, SNAI1, SNAI2, HES1, HEY1, N0TCH1, WNT5A) previously reported to inhibit myogenic differentiation through repression of MyoD activity, activation of EMT programs, or maintenance of progenitor states via TGF-P and Notch signaling pathways (Benezra et al., 1990; Spicer et al., 1996; Buas et al., 2009; Soleimani et al., 2012; von Maltzahn et al., 2012) and performed barrier score analysis that are regulatory drivers of the barrier genes in the myofibroblast cluster versus muscle cluster. To visualize the molecular landscape of reprogramming resistance, we correlated gene expression profiles with a composite " Barrier Score," representing the cellular state locked in an activated fibroblast / myofibroblast identity. Our analysis identified a hierarchical "brake" system categorized into four distinct regulatory drivers that actively antagonize myogenic transdifferentiation. The most potent barrier was the structural reinforcement of the ECM, characterized by strong positive correlations with COL1A2, SPARC, and COL3A1 suggesting the physical rigidity and integrin signaling of the matrix structurally anchor cells to a non-myogenic identity. Active myofibroblast markers, including TAG LN, CTGF, INHBA, and POSTN, showed significant positive correlation with the barrier score, confirming that a competing pro-fibrotic program directly diverts cells from the skeletal muscle fate. We identified active molecular inhibitors of myogenesis, specifically IGFBP5, IGFBP7, and ID3. The latter is a known negative regulator of basic helix-loop-helix transcription factors, such as MyoD, suggesting these factors biochemically quench the myogenic program. Regulatory genes associated with cellular stress and inflammation specifically IT' 16, STAT3, and NFKB1, positively correlated with the barrier score. Collectively, these results map a multi-layered roadblock to myogenesis, where successful reprogramming requires the simultaneous suppression of structural ECM traps, competing fibrotic lineages, transcriptional repressors, and inflammatory stress states (Fig. 6D).

[0962] To identify potent protein ligands from our synthetic library, we first sought to establish a streamlined, serum-free screening assay. Since prolonged induction, extending to 14 days of Doxycycline (Dox) treatment did not further improve conversion efficiency, we conducted a temporal screen to define the minimal Dox exposure required to prime the fibroblasts for myogenic reprogramming. We evaluated Dox treatment durations of 2, 4, 6, 8, 10, and 14 days. Following induction, cells were transitioned into serum-free media for various release periods to observe the progression of differentiation. Our time-course analysis revealed that a 4-day Dox pulse (D4) followed by a 4-day release into serum-free media (D8) was sufficient to drive consistent myogenic conversion.Under these optimized D8 conditions, approximately 30% of the population expressed the intermediate filament Desmin and Myosin heavy chain. Additionally, we observed significant metabolic remodeling, a hallmark of the transition from a fibroblastic state to a mature myogenic oxidative profile (Fig. 6E). This optimized D8 window provides a clear baseline for evaluating whether our Al-designed minibinders can surpass the -30% efficiency barrier and accelerate functional maturation.

[0963] Building on the characterization of the D8 optimized window, we investigated the metabolic shift that accompanies myogenic reprogramming. MyoD is known to bind the PGC1B promoter to drive mitochondrial biogenesis, and our data confirm a robust metabolic transition during the conversion process. We observed a significant increase in mitochondrial activity and structural maturation across the transition from fibroblast to myocyte. Desminpositive reprogrammed cells exhibited a dramatic increase in ATPB synthase expression (Fig.

[0964] 6F). Both the D4 (induction) and D8 (post-release) conditions demonstrated significantly elevated mitochondrial respiration. To quantify these functional changes, we performed a Mito Stress Test using Seahorse extracellular flux analysis at DO, D4, and D8. There was a progressive and significant increase in maximal mitochondrial respiration from DO to D8. We observed a modest increase in basal respiration, consistent with the higher energy demands of myogenic commitment (Fig. 6G). These results validate that the D8 condition in the HFF-iMyoD line not only achieves structural myogenic marker expression but also captures the essential metabolic "switch" required for functional muscle tissue. Consequently, we established this D8 timeframe as our standardized screening platform to evaluate the efficacy of Al-designed minibinders.

[0965] Designed receptor modulatory cocktail (DPC) enhance the skeletal muscle direct reprogramming

[0966] Despite its role as a master regulator, MyoD expression alone is insufficient to drive efficient myogenic reprogramming in human fibroblasts (Zhang et al., 2022). This limitation is largely attributed to epigenetic constraints, incomplete metabolic maturation, and barriers associated with cellular aging. Specifically, chromatin inaccessibility at key myogenic loci acts as a primary bottleneck, leading to incomplete transdifferentiation and a lack of functional maturity (Manandhar etal., 2017).

[0967] To date, numerous biological ligands including Follistatin, GDF8, FGF2, GDF11, GDF15, hGH, TMSB4X, BMP4, BMP7, IL-6, and TNF-a, have been evaluated for their ability to enhance this process but none of these are successful examples. However, thesefactors predominantly promote cellular proliferation or function as negative regulators that further impede myogenic conversion (Abdel-Raouf et al., 2021). In contrast, targeted modulation of signaling pathways, such as Wnt activation (CHIR99021) or Notch inhibition (DAPT), has demonstrated that specific signaling cues can improve both the efficiency and maturity of transdifferentiated muscle cells (Xu et al., 2020).

[0968] Leveraging recent advances in computational protein design, we hypothesized that precise outside-in signaling control using engineered minibinders could overcome fatestabilizing barriers and promote productive myogenic reprogramming. We therefore employed an optimized D8 reprogramming paradigm, in which minibinders were applied under serum-free conditions between day 4 and day 8, a critical window for fate stabilization (Fig. 6H). We first performed a systematic screen of individual minibinders targeting FGFR, ALK1R, PDGFR, InsulinR, IGF1R, TGFBRII, and EGFR as monomeric antagonists, together with engineered agonists including FGFR-C6, TRKA-C2, InsulinR, Tie2-H8, and EGFR (Fig. 6H, I). We analysed the desmin intensity of each condition (Fig. 61). From the screen, we identified two design protein cocktails (DPC) that significantly enhanced myogenic conversion: (I) TGFBRIImb + ALKIRmb + FGFR-C6 agonist and (II) TGFBRIImb + ALKIRmb + TRKA-C2 agonist where ALK1 and TGFBR2 pathway inhibition remains central in both the DPCs (Fig. 61). The amino acid sequences for the TGFBRIImb + ALKIRmb + FGFR-C6 agonist components are shown in Table 16.

[0969] Alkl pathway strong suppression is efficient for the myogenic transdifferentiation To validate the mechanism of the designed receptor modulatory cocktails, we focused on the inhibition of the Alkl pathway. We utilized the IGF2R_EndoTag4-Alkl novokine to test Alkl inhibition by an alternative modality (Li et al., 2025). IGF2R_EndoTag4 minibinder facilitates the endosomal internalization and subsequent lysosomal degradation of bound ligand complexes (Fig. 7A). We therefore engineered an IGF2R_EndoTag4-Alklmb fusion designed to deplete ALK1 protein from the plasma membrane. We tested two orientations of fusion proteins- Alklmb: IGF_EndoTag4 mb (AL) and IGFR2_EndoTag4 mb: Alklmb. To assess their efficacy, we treated CHO-human ALK1 cells for 24 hours and observed that both AL and LA fusions induced a significant reduction in hALKl protein levels (Fig. 7B).

[0970] ALK1R levels returned to baseline 24 hours after the removal of the fusions, confirming the specificity and reversibility of the EndoTag4 system (Fig. 7C).

[0971] We then tested the Alklmb: IGF_EndoTag4 mb (AL) in the transdifferentiation assay (Fig. 7D). Importantly, Alklmb-EndoTag treatment significantly increased both myogenictransdifferentiation efficiency and myotube length, further supporting the conclusion that repression of Alkl signaling pathway enhances myogenic transdifferentiation (Fig. 7D), The superior activity of Alklmb-EndoTag compared with Alklmb alone indicates that physical depletion of ALK1 from the cell surface lowers the reprogramming barrier more effectively than simple inhibition of its kinase activity. These findings suggest that receptor abundance itself, rather than catalytic signaling alone, constitutes a key constraint on cell fate conversion. More broadly, our results establish Al-designed minibinder-driven targeted protein degradation (TPD) as a powerful strategy to remove inhibitory signaling nodes. Using this approach, we identify ALK1R and TGFBRII as major molecular roadblocks to efficient muscle transdifferentiation and demonstrate that their programmable elimination can unlock latent myogenic potential.

[0972] Designed protein induces skeletal muscle transdifferentiation

[0973] We analyzed the two powerful design minibinder cocktails, C2 and C6. When compared to non-minibinder treated controls, myotubes treated with the C6 or C2 cocktails both exhibited several key hallmarks of advanced maturation. A higher percentage of fibroblasts successfully transitioned into the myogenic lineage, surpassing 3x over the -20% baseline observed in standard conditions (Fig. 8A). There was a measurable increase in both the length and width of the resulting myotubes, indicating more robust protein synthesis and sarcomeric assembly (Fig. 8B, C). Treated cells showed a significant increase in fusion index, characterized by the presence of multiple nuclei within a single continuous cytoplasm, a critical step for functional muscle contraction) (Fig. 8A, D). While both cocktails outperformed the control, the C6 cocktail (FGFRl / 2c agonist + ALK1R / TGFBRII antagonists) consistently produced the most mature phenotypes, aligning with our findings that FGFRl / 2c activation provides a better efficiency.

[0974] Cooperative regulation of promoter accessibility enhanced during C6 cocktail induced myogenic reprogramming

[0975] Bulk RNA sequencing revealed that treatment with the designed protein cocktail (DPC) induces a robust transcriptional program consistent with myogenic differentiation and muscle maturation in transdifferentiated skeletal muscle (tSKM) cells (Fig. 8E). DPC treatment significantly upregulated genes associated with muscle progenitor activation, cytoskeletal remodeling, and paracrine signaling, including MSX1, HGF, SVIL, FIBCD1, SNAI1, CMKLR1, PTGS2, and PGF, reflecting a coordinated shift toward a pro-myogenicand growth-permissive state. Gene Ontology enrichment analysis revealed activation of PI3K-AKT, MAPKZERK, JAK-STAT, calcium, VEGF, Wnt signaling, and arachidonic acid metabolism, while Reactome analysis highlighted enrichment of ERK / MAPK cascades, Notch signaling, and prostaglandin biosynthesis, collectively indicating enhanced growth factor responsiveness, survival signaling, and metabolic remodeling downstream of DPC treatment. Notably, induction of PTGS2 and downstream prostaglandin pathways, including prostaglandin F2, aligns with established roles of prostaglandin signaling in muscle growth and regeneration. The upregulation of HGF and the myokine FIBCD1 further suggests activation of autocrine and paracrine programs that support myofiber growth and tissue remodeling (Tatsumi et al., 1998; Graga et al., 2022), consistent with the increased myotube dimensions observed following DPC treatment. Developmental regulators such as ID3 and SNAI1, that are associated with early lineage plasticity and morphogenetic programs, were also elevated (Benezra et al., 1990; Soleimani et al., 2012), suggesting partial reactivation of developmental transcriptional states that may synergize with chromatin remodeling to enhance maturation. In addition, increased expression of CCN3 / NOV, an extracellular matrix-associated factor that cooperates with growth factor signaling, points to improved extracellular matrix interactions that further support muscle cell physiology (Lafont et al., 2005).

[0976] To identify transcriptional regulators underlying enhanced myogenic reprogramming induced by the FGFRlc-agonist based C6 protein cocktail (C6-DPC), we performed motif enrichment analysis on regulatory regions associated with genes upregulated during conversion. We extracted promoter sequences for the 185 upregulated genes by defining each promoter as a fixed window spanning -2000 bp upstream to +200 bp downstream of the transcription start site (TSS). First, the 185 genes were mapped to their corresponding genomic coordinates and strand information using a reference genome annotation (e.g., hg38 / GRCh38). Our motif enrichment analysis on regulatory regions associated with it revealed a significant enrichment of KLF family motifs, characterized by GC-rich CACCC elements, together with ETS family motifs, including the canonical GGAA sequence recognized by ETV1 and ETV4 (Fig. 8F). Treated cells showed a significant increase in fusion index, characterized by the presence of multiple nuclei within a single continuous cytoplasm, a critical step for functional muscle contraction (Fig. 8F). Notably, KLF and ETS motifs frequently co-occurred within promoter-proximal and enhancer regions of genes implicated in cytoskeletal organization, metabolic remodeling, and early myogenic commitment. We propose a model in which coordinated inhibition of TGFpR and ALK1Rsignaling leads to attenuation of SMAD2 / 3 activity, resulting in stabilization and functional activation of KLF4 which primes chromatin accessibility and binds the HMGA2 promoter. Concurrent FGFR1 mediated MAPK activation induces ETS transcription factors, including ETV1 / 4, which co-target HMGA2 regulatory regions. Upregulation of HMGA2 drives global chromatin decompaction, removing epigenetic barriers and enabling efficient MyoD / other MRFs-driven activation of the myogenic program.

[0977] C6 Cocktail enhances myogenic remodeling

[0978] Transcriptomic analysis revealed that both C2 and C6 cocktails induce MyoD-mediated fibroblast reprogramming leading to muscle marker expression (MYH7, ACTA1, TTN, MYL4, CASQ2 etc) and reduced expression of fibroblast marker (FBLN5, FN1, FGF5). Both C2 and C6 conditions further induced a myogenic reprogramming response, as demonstrated by the upregulation of skeletal muscle-associated genes (LM0D3, MYL6B, COX6A2, PDLIM4, and MYO 18 A) involved in sarcomere assembly and contractile function (Fig. 9A,)

[0979] C6-DPC treatment triggered myogenic marker SNTB1, extensive extracellular matrix (ECM) remodeling, characterized by increased expression of collagens and matrix-modifying enzymes (such as COL5A3, C0L6A1, C0L7A1, BGN, CSPG4, MMP1, MMP3, and MMP27) consistent with an activated or transitional reprogramming state (Fig. 9B).

[0980] Importantly, C6 treatment further showed evidence of metabolic maturation, marked by increased expression of genes involved in glycolysis and mitochondrial energy metabolism, including PFKL, PKM, EN01, ACADVL, ATP5F1D, COX6A2, and CKB, suggesting enhanced bioenergetic capacity accompanying myogenic conversion.

[0981] C6-DPC specifically suppresses the interferon / inflammatory barrier (IFIT1, IFIT3, IFI44L, CXCL11) but some inflammatory and NF-KB-associated genes (such as ILIA, IL1B, IL24, IL32, CSF2, CSF3, CXCL8, NFKBIA, NFKB2, STAP2, and TNFRSF21) remained significantly upregulated under C6 conditions, indicating that myogenic reprogramming occurs alongside a persistent inflammatory transcriptional program rather than complete resolution of the fibroblast state (Fig. 9B).

[0982] This productive module of C6-DPC was characterized by induction of sarcomeric and cytoskeletal assembly genes such as LM0D3, UNC45A, and SNTB1, together with mitochondrial and oxidative metabolism components including COX6A2 and ACADVL. Notably, inflammatory and ECM transcripts remained detectable, indicating that C6 does not globally suppress fibroblast or stress-associated programs. Instead, biased FGFR signalingcombined with ALK1R and TGFBR2 inhibition selectively enables myogenic structural and metabolic maturation to dominate over fate-stabilizing fibroblast programs. These findings support a model in which efficient lineage conversion is achieved not by erasing inflammatory identity, but by functionally overriding it through targeted signaling bias that unlocks irreversible myogenic commitment. To confirm the metabolic remodeling we show that C6 cocktail induced increased mitochondrial mito orange uptake indicating improved bioenergetic capacity (Fig. 9C, D). Further, prolonged C6-cocktail treated cells exhibited pronounced sarcomeric organization and maturation stained for alpha actinin (Fig. 9E). These data indicated that C6 FGFRlc-biased signaling as a dominant determinant of functional myogenic reprogramming.

[0983] Inflammation overrides promyogenic signal

[0984] Single-cell transcriptomic profiling of cells that did not commit to a myogenic fate revealed marked differences between C6-DPC treated and no mb control conditions (Fig. 9F). Pseudo-bulk analysis of the unconverted populations identified a significant induction of inflammatory pathways (Fig. 9F), such as TNFRSF19 and TNFRSF21 specifically in C6-DPC treated cells. Elevated TDO2, PLAT, and MMP expression in non-committed C6-treated cells further indicates activation of a TNF-associated, stress-adapted, ECM-remodeling transcriptional program, revealing that C6 signaling actively redirects unresponsive cells toward a distinct, non-myogenic reprogramming state rather than maintaining a fibroblast-like identity.

[0985] To investigate the impact of the inflammatory "roadblock" identified in our earlier correlation analysis, we challenged the C6-DPC (Designed Protein Cocktail) with TNF-a treatment (Fig. 9G). While the C6 cocktail typically drives high-efficiency myogenic conversion, the addition of TNF-a prevented the characteristic increase in conversion efficiency normally induced by the C6-DPC (Fig. 9G). This finding suggests that inflammatory stress acts as a dominant inhibitory checkpoint that can override the promyogenic signals provided by FGFRl / 2c activation and ALK1R / TGFBRII inhibition. High fibrosis roadblock for myogenic transdifferentiation (Fig. 9H)

[0986] Designed protein cocktail promotes iPSCs into muscle differentiation

[0987] To determine whether the C6 designed protein cocktail enhances myogenic differentiation and maturation of human pluripotent stem cell-derived muscle context, we applied the cocktail to an inducible MyoD iPSC differentiation system followed by structuraland functional analyses. WTC11 human iPSCs carrying a doxycycline-inducible MyoD cassette were treated with doxycycline for 2 days to initiate myogenic commitment, followed by 4 days of differentiation in skeletal muscle medium supplemented with fetal bovine serum (Fig. 10A). Cells were subsequently exposed to the C6 FGFRlc-agonist-based cocktail for an additional 4 days before fixation and immunostaining. Immunofluorescence analysis revealed robust formation of multinucleated myotubes expressing canonical muscle markers desmin and Myosin Heavy Chain (MHC) (Fig. 10B). Quantitative image analysis showed that C6 treatment significantly increased myotube area coverage relative to control conditions lacking minibinders (Fig. IOC). Consistent with enhanced structural maturation, the mean fluorescence intensity of MHC was significantly elevated in C6-treated cultures (Fig. 10D), indicating increased sarcomeric protein accumulation and myofiber maturation.

[0988] C6-DPC enhances structural and functional maturation of iPSC-derived engineered muscle tissues

[0989] To determine whether C6-DPC-induced improvements in myogenic differentiation translate to functional gains in a 3D tissue context, we generated engineered muscle tissues (EMTs) from both wild-type and dystrophin-deficient (DMD) iPSC-derived myoblasts and assessed contractile function using the Mantarray™ magnetometric platform (Fig. 10E). Immunofluorescence cross-sections of C6-treated EMTs revealed denser, more organized myofiber architecture compared to PBS-treated controls in both genotypes (Fig. 10F), quantified by a significant increase in effective cross-sectional area (CSA), which increased by 33% in WT (PBS: 0.229 vs C6: 0.304 mm2) and 29% in DMD EMTs (PBS: 0.128 vs C6: 0.165 mm2) (Fig. 10H). C6 treatment also increased mean myotube diameter in WT tissues (PBS: 12.24 vs C6: 15.06 pm), though this hypertrophic effect was not significant in DMD EMTs (Fig. 10G), suggesting that the increased CSA in DMD tissues reflects enhanced myoblast fusion and fiber density rather than individual fiber hypertrophy. Critically, these structural gains translated directly into enhanced contractile output. Under tetanic stimulation (100 Hz), C6-treated WT EMTs generated ~2.2 fold greater active force compared to PBS controls (PBS: 235.2 vs C6: 515.9 pN), with DMD tissues showing a comparable ~3.9 fold increase (PBS: 90.2 vs C6: 346.8 pN) (Fig. 101). Single-twitch responses (0.2 Hz) followed the same pattern, with C6 treatment increasing active twitch force by ~2.1 fold in WT and ~5.8 fold in DMD EMTs (Fig. 10 J).

[0990] However, increased absolute force could simply reflect the larger tissue cross-section rather than improved intrinsic muscle quality. To distinguish between these possibilities, wecalculated specific force by normalizing active force to effective CSA. Notably, C6-treated EMTs showed significantly elevated specific force under both tetanic and twitch stimulation in both genotypes (Fig. 10K), demonstrating that the contractile improvement exceeds what can be attributed to increased tissue size alone, indicating that C6 treatment enhances the force-generating capacity of the myofibers themselves. To further assess functional maturation, we examined relaxation kinetics following tetanic contraction. C6 treatment significantly accelerated the time to 80% force relaxation (RT80%) in both WT (PBS: 0.505s vs C6: 0.430s, A75 ms) and DMD tissues (PBS: 0.560s vs C6: 0.460s, A100 ms) (Fig. 10L).

[0991] Faster relaxation is a hallmark of mature skeletal muscle, reflecting efficient calcium reuptake and cross-bridge detachment kinetics

[0992] Collectively, these results demonstrate that C6-DPC does not simply produce larger muscle, it produces better muscle. The convergence of increased specific force, accelerated relaxation kinetics, and enhanced myofiber fusion defines a coordinated maturation program that is preserved across both healthy and dystrophin-deficient backgrounds, establishing synthetic receptor-specific signaling as a viable strategy for functional muscle regeneration.

[0993] Discussion

[0994] Direct lineage conversion holds great promise for regenerative medicine but remains limited by inefficient fate transitions and incomplete maturation of reprogrammed cells. Our study demonstrates that programmable extracellular signaling using Al-designed synthetic minibinders can overcome key barriers that restrict myogenic transdifferentiation. By defining an optimized D8 reprogramming window and applying single-cell transcriptomic analysis, we identified that the majority of fibroblasts resist conversion through a coordinated barrier network involving extracellular matrix reinforcement, fibrotic lineage commitment, and transcriptional inhibitors of myogenesis. This barrier landscape reveals that inefficient reprogramming is not a stochastic process but rather reflects a structured regulatory program that stabilizes fibroblast identity.

[0995] Targeted manipulation of this barrier network using engineered receptor modulators proved sufficient to unlock productive lineage conversion. The superior performance of the C6 cocktail combining FGFRl / 2c agonism with inhibition of ALK1 and TGFBR2 signaling, demonstrates that efficient myogenic reprogramming requires simultaneous activation of pro-myogenic pathways and suppression of fate-stabilizing signaling. Unlike natural growth factors, which often trigger pleiotropic or proliferative responses, synthetic minibinders enable receptor-isoform-specific control of signaling architecture. This precision allowsselective attenuation of fibrotic signaling programs while reinforcing metabolic and structural pathways associated with muscle differentiation. The observed increase in mitochondrial activity and ATP synthase levels further suggests that metabolic remodeling is a critical enabling step during the acquisition of functional myogenic identity.

[0996] Our findings also highlight the importance of inflammatory signaling as a dominant regulatory checkpoint. The inhibitory effect of TNF-a on C6-mediated conversion indicates that inflammatory pathways can override pro-myogenic signaling cues, providing a mechanistic explanation for the reduced regenerative capacity observed in aging and muscular dystrophy. These results suggest that modulation of inflammatory signaling may represent a complementary strategy to further improve reprogramming efficiency in diseased or aged tissues.

[0997] Importantly, the effects of C6-DPC extend beyond structural differentiation to intrinsic functional maturation. In engineered muscle tissues, C6 treatment increased not only absolute contractile force but also specific force. C6-treated cells exhibited enhanced myotube maturation and significantly improved twitch and tetanic force generation in engineered human muscle tissues, including those derived from dystrophin-deficient iPSCs. This functional rescue suggests that precise control of receptor signaling can partially compensate for the contractile deficits associated with several muscle-related disorders.

[0998] Together, these findings establish programmable synthetic ligands as a versatile platform for rewiring receptor signaling networks to control cell fate transitions. By combining pathway activation, inhibition, and targeted receptor degradation within a single design framework, Al-designed minibinders enable rational engineering of cellular signaling environments. This strategy provides a generalizable approach for overcoming lineage barriers and enhancing functional tissue regeneration, with potential applications in muscle repair, degenerative disease modeling, and precision regenerative medicine.

[0999] Example 3. Strong TGFbR2 / ALK1R inhibitor novokine

[1000] We showed using novokines novel modality that forces unrelated receptors together to induce novel signaling in Example 1. Our screen suggested that a few novokines can enhance myogenic transdifferentiation, novokines in general can have two modalities, antagonism (for example in cellular context where only one receptor is present), or novel agonism. We tested between these modalities. We also showed in Example 2 that a cocktail consisting of either TRKA C2 agonist or FGFRl / 2c agonist (C6FGFmb) in combination withtwo antagonists (Alklmb and TGFBRIImb) enhanced myogenic transdifferentiation efficiency.

[1001] Therefore, we tested the uncharacterised HFs containing TGFRIImb or ALKlmb for ALK1 or TGFP pathway inhibition respectively (Fig. 12B). In a 15 min competition assay with TGFP, we found that the HFs consisting of TGFbRIImb reduced the pSmad2 / 3 levels (Fig. 12D). TGFBRII-GP130 and TGFBRII-TGFBRII HFs turned out to be stronger inhibitors of TGFP pathways than TGFRIImb alone (Fig. 12D). Similarly, Novokines with AlKlmb showed a strong inhibition of BMP9-induced increase in pSmadl / 5 levels (Fig. 12C). IGF_EndoTag4 mb binds to the Mannose-6-receptor (M6R) where the M6R bound to ligand complex gets internalized ultimately into an acidic lysosomal compartment for enzymatic degradation (IGF_EndoTag4). To test if Alklmb: IGF_EndoTag4 mb (AL) and IGF_EndoTag4 mb: Alklmb (LA) results in Alkl lysosomal degradation, we treated CHO-hALKl cells for 24 hours with these Novokines and observed a dramatic downregulation of the hALKl receptor (Fig. 12E). This reduction was reversible for 24 hours after removal of HF (LA or AL), ALK1 levels returned to normal (Fig. 12F-H). Our data suggest that the mode of action of HFs containing TGFRIImb or ALKlmb on increased myogenic differentiation is plausible through TGFb and ALK1 pathway inhibition. We therefore continued to explore the molecular modality of TAB2 and H2F that were identified by both screens and are not predicted to act through antagonism.

[1002] Mechanism of TAB2 novokines

[1003] TRKA: BMPR2 novokines

[1004] Another uncharacterized Novokine with a distinct receptor combination, TRKA-BMPR2 (SEQ ID NO: 1464 ) (TAB2) (Fig. 13A), in which TRKA (tropomyosin receptor kinase A; NTRK1) is an RTK and BMPR2 (bone morphogenetic protein receptor type II) is an RSTK, also significantly accelerated myogenic fate acquisition in transdifferentiation assays. To test whether this effect could be generalized, we expanded our screen to additional novokines pairing multiple RSTK-targeting minibinders (ALK2, ALK3, TGFBR2, BMPR2, ACVR2A, and ACVR2B) with diverse RTK-targeting minibinders. This analysis identified several positive RSTK-RTK novokines, including BMPR2-HER2, BMPR2-EGFR, BMPR2-IGF1R, ACVR2A-TRKA, and ACVR2A-EGFR, that further enhanced myogenic transdifferentiation (Table 15). We additionally identified RSTK-RSTK novokines, such as ALK1-ALK3, ALK1-BMPR2, and ACVR2B-ALK1, that also improved conversion efficiency. Together, these findings indicate that novokine activity is not restricted toconventional RTK-RTK pairings, but can also emerge from engineered RSTK-RTK and RSTK-RSTK receptor combinations, revealing a broader and previously unappreciated design space for synthetic control of cell fate.

[1005] These results are particularly notable because RSTK-RTK Novokines represent a mechanistically unusual class of synthetic ligands that bridge two fundamentally distinct receptor signaling systems. Their activity suggests that enforced proximity between heterologous receptor classes can generate productive, noncanonical signaling states that promote lineage conversion.

[1006] We next dissected the mechanism of the TRKA-BMPR2 novokine (SEQ ID NO: 1464) (TAB2) (Fig. 13 A) and observed a robust enhancement of myogenic maturation, including increased fusion index, multinucleation, and myotube hypertrophy (Fig. 13B-E). In addition to structural maturation, TAB2 promoted metabolic remodeling, as evidenced by increased mitochondrial biogenesis and respiratory capacity, reflected by elevated ATP synthase P subunit levels and MitoTracker Orange intensity (Fig. 13F).

[1007] To define the signaling mode of TAB2, we performed phospho-effector profiling and identified strong TAB2-dependent activation of ERK and CREB pathways (Fig. 13 J). Doseresponse analysis revealed that TAB2 behaves as a high-affinity agonist, EC-50 of ~6 nM based on pERK induction in CHO cells co-expressing human BMPR2 and TRKA receptors (Fig. 13G). Notably, TAB2-mediated signaling strictly required co-expression of both receptors, as pERK activation was abolished when either TRKA or BMPR2 was expressed alone (Fig. 3H), indicating that receptor pairing is essential for signal propagation.

[1008] Consistently, competitive inhibition with either TRKA- or BMPR2-targeting minibinders significantly attenuated TAB2-induced pERK and pCREB activation (Fig. 13H, I), further confirming dual-receptor dependency.

[1009] Finally, pharmacological inhibition of TRKA kinase activity using selitrectinib or entrectinib markedly reduced TAB2-induced ERK phosphorylation (Fig. 131), demonstrating that TRKA catalytic activity is required for downstream signaling. Together, these data establish TAB2 as a bona fide agonist that enforces TRKA-BMPR2 receptor coupling to drive coordinated MAPK-CREB signaling and promote functional myogenic maturation.

Claims

We claim1. A fusion protein, comprising:(a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508 wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; and(b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-11, 16-32, 37-41, 34-135, 137-154, 156-408, 410-1448, and 1502-1508, wherein the first polypeptide binds to a target as noted in the table in which the amino acid sequence is listed; wherein the first polypeptide and the second polypeptides bind to different targets;wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

2. The fusion protein of claim 1, wherein(a) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the first polypeptide binds to Human Epidermal Growth Factor Receptor 2 (Her2); and(b) the second polypeptide comprises:(i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 or 410-1031, wherein the second polypeptide binds to Fibroblast Growth Factor Receptor (FGFR); or(b) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 39, wherein the second polypeptide binds to Transforming Growth Factor Beta Receptor 2 (TGFBR2);wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

3. The fusion protein of claim 2, wherein the first polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, and the second polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from SEQ ID NOS: 11 or 410- 1031.

4. The fusion protein of claim 3, wherein the first polypeptide comprises the amino acid selected from SEQ ID NOS: 1-10, and the second polypeptide comprises the amino acid sequence selected from SEQ ID NOS: 11 or 410- 1031.

5. The fusion protein of any one of claims 1-4, wherein the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:2.

6. The fusion protein of any one of claims 1-5, wherein the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11 or 1031.

7. The fusion protein of any one of claims 1-6, wherein the amino acid linker is present.

8. The fusion protein of claim 7, wherein the amino acid linker comprises a GS-rich amino acid linker.

9. The fusion protein of claim 8, wherein the GS-rich amino acid linker comprises (GSGSGSGSGS)n, (SEQ ID NO: 1509) wherein n = 1-7.

10. The fusion protein of any one of claims 1-9, comprising the amino acid sequence selected from SEQ ID NOS: 12-15 and 1457.

11. The fusion protein of claim 1, comprising:(a) a first polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, wherein the first polypeptide binds to Tropomyosin receptor kinase A (TrkA); and(b) a second polypeptide comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NOS:31 or 32, wherein the second polypeptide binds to Bone Morphogenetic Protein Receptor Type 2 (BMPR2);wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker.

12. The fusion protein of claim 11, wherein the first polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NOS:31 or 32.

13. The fusion protein of claim 11, wherein the first polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NOS: 16-30, and the second polypeptide comprises the amino acid sequence of SEQ ID NOS:31 or 32.

14. The fusion protein of claim 11, wherein the first polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16, and the second polypeptide comprises an amino acid sequence at least at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:31.

15. The fusion protein of any one of claims 11-14, wherein the amino acid linker is present.

16. The fusion protein of claim 15, wherein the amino acid linker comprises a GS-rich amino acid linker.

17. The fusion protein of claim 16, wherein the GS-rich amino acid linker comprises (GGSGGSGGSG)n (SEQ ID NO: 1478), wherein n = 1-718. The fusion protein of any one of claims 11-17, comprising an amino acid sequence selected from SEQ ID NOS:33-36.

19. The fusion protein of claim 1, wherein:(a) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38, wherein the first polypeptide binds to Activin receptor-like kinase 1 (Alkl); and(b) the second polypeptide comprises(i) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1-10, wherein the second polypeptide binds to HER2; or (ii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357:, wherein the second polypeptide binds to Epidermal Growth Factor Receptor (EGFR); or(iii) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359, wherein the second polypeptide binds to interleukin-2 receptor subunit gamma (IL-2RG); or(iv) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1360-1448, wherein the second polypeptide binds to IL-2 receptor βγc heterodimer; or(v) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408, wherein the second polypeptide binds to insulin receptor; or(vi) an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508, wherein the second polypeptide binds to Insulin-like Growth Factor Receptor 2 (IGFR2);wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.

20. The fusion protein of claim 1, wherein:(a) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence SEQ ID NO:39that bind to Transforming Growth Factor Beta Receptor 2 (TGFBR2); or(b) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to GammaC, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to Platelet-Derived Growth Factor Receptor (PDGFR); or(c) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to interleukin-2 receptor subunit gamma (IL-2RG), and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2; or(d) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2 and, the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1036-1335that bind to PDGFR; or(e) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 99-135, 137-154, and 156-408that bind to insulin receptor, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG; or(f) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acidsequence selected from SEQ ID NO:39that bind to TGFBR, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1032-1035that bind to Gpl30; or(g) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30that bind to TrkA, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2; or(h) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30that bind to TrkA; or(i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 31-32that bind to BMPR2; or(j) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1358-1359 that bind to IL-2RG, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39 that bind to TGFBR; or(k) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508that bind to IGFR2, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 and 410-103 Ithat bind to FGFR; or(l) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1360-1448that bind to IL-2 receptor βγc heterodimer, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1336-1357that bind to EGFR; or(m) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NO:39that bind to TGFBR, and the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 40-51, 43-125, and 1508thatbind to IGFR2;wherein the first polypeptide and the second polypeptide are optionally connected by an amino acid linker, and where the first and second polypeptide can be in any order in the fusion protein.

21. The fusion protein of claim 19 or 20, wherein the first polypeptide and the second polypeptide comprise an amino acid sequence at least 80% identical to the reference sequence.

22. The fusion protein of claim 19 or 20, wherein the first polypeptide and the second polypeptide comprise an amino acid sequence at least 90% identical to the reference sequence.

23. The fusion protein of claim 19 or 20, wherein the first polypeptide and the second polypeptide comprise an amino acid sequence at least 95% identical to the reference sequence.

24. The fusion protein of any one of claims 19-23, wherein the amino acid linker is present.

25. The fusion protein of claim 24, wherein the amino acid linker comprises a GS-rich amino acid linker.

26. The fusion protein of claim 25, wherein the GS-rich amino acid linker comprises (GSGSGSGSGS)n (SEQ ID NO:1509)wherein n = 1-7.

27. The fusion protein of any one of claims 1-26, comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500, wherein optional residues may be present or may be deleted in whole or in part.

28. A fusion protein, comprising an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 1449-1475 and 1479-1500.

29. A nucleic acid encoding the fusion protein of any one of claims 1-28.

30. An expression vector comprising the nucleic acid of claim 29 operatively linked to a suitable control sequence, such as a promoter.

31. A host cell comprising the fusion protein, nucleic acid, or expression vector of any one of claims 1-30.

32. A composition, comprising at least a first and a second polypeptide according to any preceding claim, wherein the first and second polypeptides bind to different targets, and wherein the first and second polypeptides are not covalently linked.

33. The composition of claim 32, wherein the composition comprises at least a first polypeptide, a second polypeptide, and a third polypeptide, wherein:(a) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 11 and 410-1031 that bind to FGFR, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl; or(b) (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS: 16-30 that bind to TrkA, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 that binds to TGFBR, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from SEQ ID NOS:37-38 that bind to Alkl.

34. The composition of claim 32, wherein the (i) the first polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11 or 1476;, (ii) the second polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:39 or 1477, and (iii) the third polypeptide comprises an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:37.

35. A pharmaceutical composition, comprising:(a) the fusion protein, nucleic acid, expression vector, host cell, and / or composition of any one of claims 1-34; and(b) a pharmaceutically acceptable carrier.

36. A method for conversion of fibroblasts to the myogenic fate, comprising contacting fibroblasts with an amount effective of the fusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to convert the of fibroblasts to the myogenic fate.

37. A method treating or limiting development of a disorder selected from muscle atrophy post-injury and muscle degeneration disorders (e.g. muscular dystrophy and sarcopenia), comprising administering to a subject in need thereof an amount effective of thefusion protein, nucleic acid, expression vector, host cell, or pharmaceutical composition of any embodiment or combination of embodiments herein to treat or limit development of the disorder.

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