Bispecific antibodies binding to FZD4 and GPR124 and / or reck

Abstract

The current disclosure relates to a soluble antibody or antibody fragment, wherein the antibody or antibody fragment is a modulator of Wnt7 signaling pathway, preferably an agonist of the Wnt7 pathway, and wherein said antibody or antibody fragment comprises one or more Fzd binding domains, and one or more Gpr124 or Reck binding domains. The disclosure also relates to uses of said antibody or antibody fragment.

Description

[0001] WNT7 SIGNALING MODULATORS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to modulators of the Wnt7 signaling pathway, more particularly to bispecific or trispecific binding agents, even more particularly bispecific or trispecific antibodies or antibody fragments, that target two or three members of the Wnt7 receptor complex. The antibodies or antibody fragments are developed to be deployed in medical treatments, such as in the treatment of neurological and / or ophthalmological diseases and / or disorders.

[0004] BACKGROUND

[0005] Endothelial Wnt / 0-catenin signaling acts as a master regulator of Blood-Brain Barrier (BBB) and Blood-Retina Barrier (BRB) physiology in response to neural-derived Wnt7a / b ligands. Wnt / 0-catenin signaling initiates the BBB and BRB differentiation cascade at the earliest steps of central nervous system (CNS) and retinal vascular invasion, and then maintains BBB and BRB function in adults. Recent evidence suggests that inhibition of Wnt signaling by conditional deletion of 0-catenin signaling in cerebral and retinal endothelial cells (ECs) causes BBB and BRB breakdown and accelerates disease progression in stroke, glioblastoma, and multiple sclerosis murine models, as well as in retinal degenerative diseases. Conversely, recombining a constitutively active form of 0-catenin in the CNS or retinal endothelium is protective in models of brain cancer, stroke, and retinal vascular dysfunction.

[0006] Wnt7a / b being the endogenous ligands controlling p-catenin-dependent BBB and BRB maturation, they are in principle, legitimate therapeutic agents to repair the dysfunctional BBB or BRB. However, activation of Wnt signaling via Wnt7 ligands has pleiotropic outcomes across a range of tissues and organs, in both health and disease. More so, the structural modalities of Wnt / Frizzled (FZD) interactions disqualify natural Wnt ligands as safe therapeutics. Interestingly, Wnt7 ligands activate Wnt signaling via two distinct types of membrane receptor complexes. The first receptor complex consisting of a member of the Frizzled (FZD) family receptors and LRP5 or LRP6 has broad tissue distribution, binds non-discriminately to Wnt7a / b and leads to systemic activation of Wnt signaling. The second one is the GPR124 / RECK / Fzd / LRP5 or LRP6 complex which is enriched at cerebral and retinal endothelial cells where GPR124 and / or RECK are expressed.

[0007] The observation that Wnt signalling at the BBB, BRB, and CNS requires a different receptor complex compared to other cell types and tissues, opens the opportunity todevelop CNS and retina specific Wnt agonists. However, developing chemical or biologicals that bind to and stimulate a four-membered receptor complex is challenging. W02019 / 180204 for instance suggests many compounds but fails to disclose concrete data or experimental evidence demonstrating the availability of such molecules or that such molecules are effective in modulating Wnt signaling specifically in the CNS or retina. The mere suggestion without enabling disclosure does not provide a viable pathway for developing such therapeutic agents.

[0008] The present invention provides a solution based on the unexpected observation that binding of a compound to only two or three specific members of the receptor complex is sufficient to activate the atypical GPR124 / RECK / FZD / LRP5 or LRP6 mediated Wnt signalling in both the BBB and BRB.

[0009] SUMMARY OF THE INVENTION

[0010] The present invention and embodiments thereof serve to provide a solution to one or more of above-mentioned disadvantages. To this end, the present invention relates to a soluble antibody or antibody fragment, wherein the antibody or antibody fragment is a modulator of Wnt7 signaling pathway according to claim 1.

[0011] Said antibody or antibody fragment is preferably an agonist of the Wnt7 pathway, and comprises one or more Fzd binding domains, and further one or more Gprl24 and / or Reck binding domains.

[0012] Preferred embodiments of the antibody or antibody fragment are shown in any of the claims 2 to 21.

[0013] In embodiments, said antibody or antibody fragment is bispecific or is trispecific. In embodiments, said antibody or antibody fragment comprises one or more Fzd binding domains and one or more Gprl24 binding domains. In embodiments, said antibody or antibody fragment comprises one or more Fzd binding domains and one or more Reck binding domains. In embodiments, said antibody or antibody fragment comprises one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains. In embodiments, said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains and Gprl24 and / or Reck binding domains.In a second aspect, the present invention relates a to nucleic acid encoding the antibody or antibody fragment of the invention or a vector comprising said nucleic acid, according to claim 22.

[0014] In another aspect, the present invention relates a to pharmaceutical composition comprising the antibody or antibody fragment, the nucleic acid, or the vector, according to claim 23.

[0015] In other aspects, the present invention relates to the use of the antibody or antibody fragment, the nucleic acid, or the vector, as a medicament according to claim 24, in blood brain barrier related conditions and / or in the treatment of neurological disorders according to claims 25-26, in blood retina barrier related conditions and / or in the treatment of ophthalmological diseases or disorders according to claims 27-28.

[0016] DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention concerns novel means and method of agonizing or stimulating the Wnt7 signaling pathway, more particularly the atypical Wnt7 signaling pathway that is mediated or controlled by the four member receptor complex comprising Fzd, LR.P5 or LR.P6, Gprl24 and Reck. Even more particularly, the modulators, for instance agonists, herein provided are specific or selective for the Wnt7 signaling pathway that is orchestrated by the GPR124 / RECK / FZD / LRP5 / 6 complex. The binding agents herein disclosed do not significantly stimulate the canonical Wnt7 signaling mediated by Fzd / LRP5 or Fzd / LRP6 alone.

[0018] Definitions

[0019] Unless otherwise defined, all terms used in disclosing the invention, including technical and scientific terms, have the meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. By means of further guidance, term definitions are included to better appreciate the teaching of the present invention.

[0020] As used herein, the following terms have the following meanings:

[0021] "A", "an", and "the" as used herein refers to both singular and plural referents unless the context clearly dictates otherwise. By way of example, "a compartment" refers to one or more than one compartment."About" as used herein referring to a measurable value such as a parameter, an amount, a temporal duration, and the like, is meant to encompass variations of + / -20% or less, preferably + / -10% or less, more preferably + / -5% or less, even more preferably + / -1% or less, and still more preferably + / -0.1% or less of and from the specified value, in so far such variations are appropriate to perform in the disclosed invention. However, it is to be understood that the value to which the modifier "about" refers is itself also specifically disclosed.

[0022] "Comprise", "comprising", and "comprises" and "comprised of" as used herein are synonymous with "include", "including", "includes" or "contain", "containing", "contains" and are inclusive or open-ended terms that specifies the presence of what follows e.g. component and do not exclude or preclude the presence of additional, non-recited components, features, element, members, steps, known in the art or disclosed therein.

[0023] Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order, unless specified. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.

[0024] The recitation of numerical ranges by endpoints includes all numbers and fractions subsumed within that range, as well as the recited endpoints.

[0025] The expression "% by weight", "weight percent", "%wt" or "wt%", here and throughout the description unless otherwise defined, refers to the relative weight of the respective component based on the overall weight of the formulation.

[0026] Whereas the terms "one or more" or "at least one", such as one or more or at least one member(s) of a group of members, is clear per se, by means of further exemplification, the term encompasses inter alia a reference to any one of said members, or to any two or more of said members, such as, e.g., any >3, >4, >5, >6 or >7 etc. of said members, and up to all said members.

[0027] Unless otherwise defined, all terms used in disclosing the invention, including technical and scientific terms, have the meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. By means of further guidance,definitions for the terms used in the description are included to better appreciate the teaching of the present invention. The terms or definitions used herein are provided solely to aid in the understanding of the invention.

[0028] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment, but may. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as would be apparent to a person skilled in the art from this disclosure, in one or more embodiments. Furthermore, while some embodiments described herein include some but not other features included in other embodiments, combinations of features of different embodiments are meant to be within the scope of the invention, and form different embodiments, as would be understood by those in the art. For example, in the following claims, any of the claimed embodiments can be used in any combination. Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. The recitation of numerical ranges by endpoints includes all numbers and fractions subsumed within that range, as well as the recited endpoints. Unless otherwise indicated, nucleotide sequences are written left to right in 5' to 3' orientation. Amino acid sequences are written left to right in amino to carboxy orientation. The headings provided herein are not limitations of the various aspects of the disclosure, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification in its entirety.

[0029] The term "antibody" as used herein, refers to an immunoglobulin (Ig) molecule or a molecule comprising an immunoglobulin (Ig) domain, which specifically binds with an antigen. "Antibodies" can be intact immunoglobulins derived from natural sources or from recombinant sources and can be immunoreactive portions of intact immunoglobulins. The term "immunoglobulin (Ig) domain" as used herein refers to a globular region of an antibody chain, or to a polypeptide that essentially consists of such a globular region. Immunoglobulin domains are characterized in that they retain the immunoglobulin fold (Ig fold as named herein) characteristic of antibody molecules, which consists of a two-layer sandwich of about seven to nine antiparallel P-strands arranged in two p-sheets, optionally stabilized by a conserved disulphide bond. The term "immunoglobulin (Ig) domain", includes "immunoglobulin constantdomain", and "immunoglobulin variable domain" (abbreviated as "IVD"), wherein the latter means an immunoglobulin domain essentially consisting of four "framework regions" which are referred to in the art and herein below as "framework region 1" or "FR1"; as "framework region 2" or "FR2"; as "framework region 3" or "FR3"; and as "framework region 4" or "FR4", respectively; which framework regions are interrupted by three "complementarity determining regions" or "CDRs", which are referred to in the art and herein below as "complementarity determining region 1" or "CDR1"; as "complementarity determining region 2" or "CDR2"; and as "complementarity determining region 3" or "CDR3", respectively. Thus, the general structure or sequence of an immunoglobulin variable domain can be indicated as follows: FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4. It is the immunoglobulin variable domain(s) (IVDs) that confer specificity to an antibody for the antigen by carrying the antigen-binding site.

[0030] The term "antibody fragment" refers to a portion of any antibody or antibody-like structure that by itself has a high affinity for an antigenic determinant or epitope, and contains one or more CDRs accounting for such specificity. In some particular embodiments, at least one of the targeting moieties of the antibody or antibody fragment of the application is a single-domain antibody, an immunoglobulin single variable domain, a heavy-chain-only antibody (VHH), a single-chain antibody (scFv), a shark heavy-chain-only antibody (VNAR), a microprotein (cysteine knot protein, knottin), a DARPin, a Tetranectin, an Affibody, an Affimer, a Transbody, an Anticalin, an AdNectin, an Affilin, a Microbody, a peptide aptamer, an alterases, a plastic antibodies, a phylomer, a stradobodies, a maxibodies, an evibody, a fynomer, an armadillo repeat protein, a Kunitz domain, an avimer, an atrimer, a probody, an immunobody, a triomab, a troybody, a pepbody, a vaccibody, a UniBody, a DuoBody, a Fv, a Fab, a Fab', a F(ab')2, a peptide mimetic molecule, or a synthetic molecule, as described in US Patent Nos. or Patent Publication Nos. US 7,417,130, US 2004 / 132094, US 5,831,012, US 2004 / 023334, US 7,250,297, US 6,818,418, US 2004 / 209243, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004 / 146938, US 2004 / 157209, US 6,994,982, US 6,794,144, US 2010 / 239633, US 7,803,907, US 2010 / 119446, and / or US 7,166,697, the contents of which are hereby incorporated by reference in their entireties. See also Storz 2011 MAbs 3: 310-317.

[0031] An "immunoglobulin domain" of this application also includes "immunoglobulin single variable domains" (abbreviated as "ISVD”), equivalent to the term "single variable domains", and defines molecules wherein the antigen binding site is present on, and formed by, a single immunoglobulin domain. This sets immunoglobulin singlevariable domains apart from "conventional" immunoglobulins or their fragments, wherein two immunoglobulin domains, in particular two variable domains, interact to form an antigen binding site. Typically, in conventional immunoglobulins, a heavy chain variable domain (VH) and a light chain variable domain (VL) interact to form an antigen binding site. In this case, the complementarity determining regions (CDRs) of both VH and VL will contribute to the antigen binding site, i.e. a total of 6 CDRs will be involved in antigen binding site formation. In view of the above definition, the antigen-binding domain of a conventional 4-chain antibody (such as an IgG, IgM, IgA, IgD or IgE molecule; known in the art) or of a Fab fragment, a F(ab')2 fragment, an Fv fragment such as a disulphide linked Fv or a scFv fragment, or a diabody (all known in the art) derived from such conventional 4-chain antibody, would normally not be regarded as an immunoglobulin single variable domain, as, in these cases, binding to the respective epitope of an antigen would normally not occur by one (single) immunoglobulin domain but by a pair of (associated) immunoglobulin domains such as light and heavy chain variable domains, i.e., by a VH-VL pair of immunoglobulin domains, which jointly bind to an epitope of the respective antigen. In contrast, immunoglobulin single variable domains are capable of specifically binding to an epitope of the antigen without pairing with an additional immunoglobulin variable domain. The binding site of an immunoglobulin single variable domain is formed by a single VH / VHH or VL domain. Hence, the antigen binding site of an immunoglobulin single variable domain is formed by no more than three CDRs. As such, the single variable domain may be a light chain variable domain sequence (e.g., a VL-sequence) or a suitable fragment thereof; or a heavy chain variable domain sequence (e.g., a VH-sequence or VHH sequence) or a suitable fragment thereof; as long as it is capable of forming a single antigen binding unit (i.e., a functional antigen binding unit that essentially consists of the single variable domain, such that the single antigen binding domain does not need to interact with another variable domain to form a functional antigen binding unit). In one embodiment of the invention, the immunoglobulin single variable domains are heavy chain variable domain sequences (e.g., a VH-sequence); more specifically, the immunoglobulin single variable domains can be heavy chain variable domain sequences that are derived from a conventional four-chain antibody or heavy chain variable domain sequences that are derived from a heavy chain antibody. For example, the immunoglobulin single variable domain may be a (single) domain antibody (or an amino acid sequence that is suitable for use as a (single) domain antibody), a "dAb" or dAb (or an amino acid sequence that is suitable for use as a dAb) or a Nanobody (as defined herein, and including but not limited to a VHH); other single variable domains, or any suitable fragment of any one thereof. Inparticular, the immunoglobulin single variable domain may be a Nanobody (as defined herein) ora suitable fragment thereof. Note: Nanobody®, Nanobodies® and Nanoclone® are registered trademarks of Ablynx N.V. For a general description of Nanobodies, reference is made to the further description below, as well as to the prior art cited herein, such as e.g. described in W02008 / 020079.

[0032] Immunoglobulin domains herein also include "VHH domains", also known as VHHs, VHH domains, VHH antibody fragments, and VHH antibodies, have originally been described as the antigen-binding immunoglobulin (Ig) (variable) domain of "heavy chain antibodies" (i.e., of "antibodies devoid of light chains"; Hamers-Casterman et al (1993) Nature 363: 446-448). The term "VHH domain" has been chosen to distinguish these variable domains from the heavy chain variable domains that are present in conventional 4-chain antibodies (which are referred to herein as "VH domains") and from the light chain variable domains that are present in conventional 4-chain antibodies (which are referred to herein as "VL domains"). For a further description of VHHs and Nanobody, reference is made to the review article by Muyldermans (Reviews in Molecular Biotechnology 74: 277-302, 2001), as well as to the following patent applications, which are mentioned as general background art: WO 94 / 04678, WO 95 / 04079 and WO 96 / 34103 of the Vrije Universiteit Brussel; WO 94 / 25591, WO 99 / 37681, WO 00 / 40968, WO 00 / 43507, WO 00 / 65057, WO 01 / 40310, WO 01 / 44301, EP 1134231 and WO 02 / 48193 of Unilever; WO 97 / 49805, WO 01 / 21817, WO 03 / 035694, WO 03 / 054016 and WO 03 / 055527 of the Vlaams Instituut voor Biotechnologie (VIB); WO 03 / 050531 of Algonomics N.V. and Ablynx N.V.; WO 01 / 90190 by the National Research Council of Canada; WO 03 / 025020 by the Institute of Antibodies; as well as WO 04 / 041867, WO 04 / 041862, WO 04 / 041865, WO 04 / 041863, WO 04 / 062551, WO 05 / 044858, WO 06 / 40153, WO 06 / 079372, WO 06 / 122786, WO 06 / 122787 and WO 06 / 122825, by Ablynx N.V. and the further published patent applications by Ablynx N.V. As described in these references, Nanobody (in particular VHH sequences and partially humanized Nanobody) can in particular be characterized by the presence of one or more "Hallmark residues" in one or more of the framework sequences. A further description of the Nanobody, including humanization and / or camelization of Nanobody, as well as other modifications, parts or fragments, derivatives or "Nanobody fusions", multivalent constructs (including some non-limiting examples of linker sequences) and different modifications to increase the half-life of the Nanobody and their preparations can be found e.g. in WO 08 / 101985 and WO 08 / 142164.

[0033] "Domain antibodies", also known as "Dabs", "Domain Antibodies", and "dAbs" (the terms "Domain Antibodies" and "dAbs" being used as trademarks by theGlaxoSmithKline group of companies) have been described in e.g., EP 0368684, Ward et al. (Nature 341: 544-546, 1989), Holt et al. (Tends in Biotechnology 21: 484-490, 2003) and WO 03 / 002609 as well as for example WO 04 / 068820, WO 06 / 030220, WO 06 / 003388 and other published patent applications of Domantis Ltd. Domain antibodies essentially correspond to the VH or VL domains of non-camelid mammalians, in particular human 4-chain antibodies. In order to bind an epitope as a single antigen binding domain, i.e., without being paired with a VL or VH domain, respectively, specific selection for such antigen binding properties is required, e.g. by using libraries of human single VH or VL domain sequences. Domain antibodies have, like VHHs, a molecular weight of approximately 13 to approximately 16 kDa and, if derived from fully human sequences, do not require humanization for e.g. therapeutical use in humans. It should also be noted that single variable domains can be derived from certain species of shark (for example, the so-called "IgNAR domains", see for example WO 05 / 18629).

[0034] Immunoglobulin single variable domains such as Domain antibodies and Nanobody (including VHH domains and humanized VHH domains), represent in vivo matured macromolecules upon their production, but can be further subjected to affinity maturation by introducing one or more alterations in the amino acid sequence of one or more CDRs, which alterations result in an improved affinity of the resulting immunoglobulin single variable domain for its respective antigen, as compared to the respective parent molecule. Affinity-matured immunoglobulin single variable domain molecules of the invention may be prepared by methods known in the art, for example, as described by Marks et al. (Biotechnology 10:779-783, 1992), Barbas et al. (Proc. Nat. Acad. Sci, USA 91: 3809-3813, 1994), Shier et al. (Gene 169: 147-155, 1995), Yelton et al. (Immunol. 155: 1994-2004, 1995), Jackson et al. (J. Immunol. 154: 3310-9, 1995), Hawkins et al. (J. Mol. Biol. 226: 889 896, 1992), Johnson and Hawkins (Affinity maturation of antibodies using phage display, Oxford University Press, 1996). The process of designing / selecting and / or preparing a polypeptide, starting from an immunoglobulin single variable domain such as a Domain antibody or a Nanobody, is also referred to herein as "formatting" said immunoglobulin single variable domain; and an immunoglobulin single variable domain that is made part of a polypeptide is said to be "formatted" or to be "in the format of" said polypeptide. Examples of ways in which an immunoglobulin single variable domain can be formatted and examples of such formats for instance to avoid glycosylation will be clear to the skilled person based on the disclosure herein.

[0035] Immunoglobulin single variable domains such as Domain antibodies and Nanobody (including VHH domains) can be subjected to humanization, i.e. increase the degreeof sequence identity with the closest human germline sequence. In particular, humanized immunoglobulin single variable domains, such as Nanobody (including VHH domains) may be immunoglobulin single variable domains that are as generally defined for in the previous paragraphs, but in which at least one amino acid residue is present (and in particular, at least one framework residue) that is and / or that corresponds to a humanizing substitution (as defined herein). Potentially useful humanizing substitutions can be ascertained by comparing the sequence of the framework regions of a naturally occurring VHH sequence with the corresponding framework sequence of one or more closely related human VH sequences, after which one or more of the potentially useful humanizing substitutions (or combinations thereof) thus determined can be introduced into said VHH sequence (in any manner known per se, as further described herein) and the resulting humanized VHH sequences can be tested for affinity for the target, for stability, for ease and level of expression, and / or for other desired properties. In this way, by means of a limited degree of trial and error, other suitable humanizing substitutions (or suitable combinations thereof) can be determined by the skilled person. Also, based on what is described before, (the framework regions of) an immunoglobulin single variable domain, such as a Nanobody (including VHH domains) may be partially humanized or fully humanized. It should be noted that the immunoglobulin single variable domains, as well as the antigen-binding chimeric protein of the invention in their broadest sense are not limited to a specific biological source or to a specific method of preparation. For example but without the purpose of being limiting, the immunoglobulin single variable domains, in particular the antigenbinding chimeric proteins of the invention, can generally be obtained: (1) by isolating the VHH domain of a naturally occurring heavy chain antibody, and further engineering of the sequence to obtain the antigen-binding chimeric protein; (2) by expression of a nucleotide sequence encoding a naturally occurring VHH domain, in a format fused to said scaffold protein of the antigen-binding chimeric protein; (3) by "humanization" of a naturally occurring VHH domain and / or scaffold protein or by expression of a nucleic acid encoding a such humanized VHH domain and / or scaffold protein, and / or antigen-binding chimeric protein; (4) by "mutation" of a naturally occurring VHH domain to reduce binding to pre-existing antibodies or by engineering of the scaffold protein fusion sites to obtain an antigen-binding chimeric protein of the invention with reduced binding to pre-existing antibodies as compared to the natural VHH; or (5) by using synthetic or semisynthetic techniques for preparing proteins, polypeptides or other amino acid sequences known per se.An "epitope", as used herein, refers to an antigenic determinant of a polypeptide, constituting a binding site or binding pocket on a target molecule (e.g. a protein to which an immunoglobulin or part thereof, antibody, VHH or ISVD is binding). "Binding" means any interaction, be it direct or indirect. A direct interaction implies a contact (e.g. physical or chemical) between two binding partners. An indirect interaction means any interaction whereby the interaction partners interact in a complex of more than two molecules. An interaction can be completely indirect (e.g. two molecules are part of the same complex with the help of one or more bridging molecules but don't bind in the absence of the bridging molecule(s)). An interaction may be partly direct or partly indirect: there is still a direct contact between two interaction partners, but such contact is e.g. not stable, and is stabilized by the interaction with one or more additional molecules. The term "binding pocket", "binding domain" or "binding site" refers to a region of a molecule or molecular complex, that, as a result of its shape and charge, associates with another chemical entity, compound, protein, peptide, antibody, single domain antibody or ISVD or VHH.

[0036] An epitope could comprise 1, 2 or 3 amino acids in a spatial conformation, which is unique to the epitope. Generally, an epitope consists of at least 4, 5, 6, 7 such amino acids, and more usually, consists of at least 8, 9, 10 such amino acids. Methods of determining the spatial conformation of amino acids are known in the art, and include, for example, X-ray crystallography and multi-dimensional nuclear magnetic resonance. A "conformational epitope", as used herein, refers to an epitope comprising amino acids in a spatial conformation that is unique to a folded 3-dimensional conformation of a polypeptide. Generally, a conformational epitope consists of amino acids that are discontinuous in the linear sequence but that come together in the folded structure of the protein. However, a conformational epitope may also consist of a linear sequence of amino acids that adopts a conformation that is unique to a folded 3-dimensional conformation of the polypeptide (and not present in a denatured state). In protein complexes, conformational epitopes consist of amino acids that are discontinuous in the linear sequences of one or more polypeptides that come together upon folding of the different folded polypeptides and their association in a unique quaternary structure. Similarly, conformational epitopes may here also consist of a linear sequence of amino acids of one or more polypeptides that come together and adopt a conformation that is unique to the quaternary structure. The term "conformation" or "conformational state" of a protein refers generally to the range of structures that a protein may adopt at any instant in time. One of skill in the art will recognize that determinants of conformation or conformational state include a protein's primary structure as reflected in a protein'samino acid sequence (including modified amino acids) and the environment surrounding the protein. The conformation or conformational state of a protein also relates to structural features such as protein secondary structures (e.g., o-helix, [3-sheet, among others), tertiary structure (e.g., the 3-dimensional folding of a polypeptide chain), and quaternary structure (e.g., interactions of a polypeptide chain with other protein subunits). Posttranslational and other modifications to a polypeptide chain such as ligand binding, phosphorylation, sulfation, glycosylation, or attachments of hydrophobic groups, among others, can influence the conformation of a protein. Furthermore, environmental factors, such as pH, salt concentration, ionic strength, and osmolality of the surrounding solution, and interaction with other proteins and co-factors, among others, can affect protein conformation. The conformational state of a protein may be determined by either functional assay for activity or binding to another molecule or by means of physical methods such as X-ray crystallography, NMR, or spin labelling, among other methods. For a general discussion of protein conformation and conformational states, one is referred to Cantor and Schimmel, Biophysical Chemistry, Part I: The Conformation of Biological. Macromolecules, W.H. Freeman and Company, 1980, and Creighton, Proteins: Structures and Molecular Properties, W.H. Freeman and Company, 1993.

[0037] A "paratope" as used herein refers to the antigen-binding site and is the part of an antibody which recognizes and binds to an antigen. The paratope of the bispecific binding agent of current disclosure thus comprises the amino acid residues of the binding agent that bind the epitope of the LR.P5 or LR.P6 protein and the amino acid residues of the binding agent that bind the epitope of the Gprl24 or Reck protein.

[0038] Antibodies or binding agents in general can bind one or more epitopes of one or more targets.

[0039] "Bispecific" as used herein in the context of current disclosure means that the binding agent can bind two different epitopes or bind two different antigens simultaneously. "Multispecific" as used herein then refers to the ability of binding two or more different binding sites or antigens simultaneously. Multispecific binding agents thus include bispecific antibodies, but it can also refer to antibodies with the ability to bind to three or more targets.

[0040] Independent of the number of targets that can be bound by the binding agent, the binding agent can comprise multiple binding sites. The number of binding sites is referred to as the valency of the antibody. A "bivalent" antibody is a typical antibody structure with two binding sites. The common structure of natural antibodies comprises two identical binding sites that bind two identical antigenic determinantsor epitopes. However, a bivalent antibody can also bind two different epitopes or two different targets. A "tetravalent" antibody is an antibody that has four binding sites. It can bind to four identical or four different antigenic determinants or epitopes, or it can bind two different epitopes or two different targets that are each bound by two binding sites.

[0041] The term "affinity" generally refers to the degree to which an antibody or other binding protein (as defined further herein) binds to a target protein so as to shift the equilibrium of target protein and binding protein toward the presence of a complex formed by their binding. Thus, for example, where an antibody and an antigen are combined in relatively equal concentration, an antibody of high affinity will bind to the antigen so as to shift the equilibrium toward high concentration of the resulting complex. The equilibrium dissociation constant KD (or KD) is commonly used to describe the affinity between a ligand and a target protein, or an antibody and its antigen. KD is the calculated ratio of koff / kon, between the antibody and its antigen and thus measures the propensity of a complex to fall apart into its component molecules. The association constant (konor kon) is used to characterize how quickly the antibody binds to its target. The dissociation constant (kOff or koff, also referred to as kdis, Kdis, Kd or kd) is used to measure how quickly an antibody dissociates from its target and is expressed as number of units that dissociated from a target per second. Hence, the lower koff is, the higher the affinity towards the target, koff and thus also KD is inversely related to affinity. A high affinity interaction is characterized by a low KD, a fast recognizing (high kon) and a strong stability of formed complexes (low koff). It will be appreciated that within the scope of the present application, the term "affinity" is used in the context of the antibody or antibody fragment that binds an epitope of the LR.P5, LR.P6, Gprl24 and / or Reck protein, more particularly the antibody or antibody fragment is "functional" in binding its target via the CDR regions of its immunoglobulin (Ig) domain.

[0042] "Amino acids" as used herein refer to the structural units (monomers) that make up proteins. They join together to form short polymer chains called peptides or longer chains called either polypeptides or proteins. These chains are linear and unbranched, with each amino acid residue within the chain attached to two neighboring amino acids. Twenty amino acids encoded by the universal genetic code are naturally incorporated into polypeptides and are called proteinogenic or natural amino acids. Natural amino acids or naturally occurring amino acids are glycine (Gly or G), Alanine (Ala or A), Valine (Vai or V), Leucine (Leu or L), Isoleucine (He or I), Methionine (Met or M), Proline (Pro or P), Phenylalanine (Phe or F), Tryptophan (Trpor W), Serine (Ser or S), Threonine (Thr or T), Asparagine (Asn or N), Glutamine (Gin or Q), Tyrosine (Tyr or Y), Cysteine (Cys or C), Lysine (Lys or K), Arginine (Arg or R), Histidine (His or H), Aspartic Acid (Asp or D) and Glutamic Acid (Glu or E).

[0043] As used herein, the terms "nucleic acid", "nucleic acid sequence" or "nucleic acid molecule" are used interchangeably and refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. Nucleic acids may have any three-dimensional structure, and may perform any function, known or unknown. Non-limiting examples of nucleic acids include a gene, a gene fragment, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, control regions, isolated RNA of any sequence, nucleic acid probes, and primers. The nucleic acid molecule may be linear or circular. The nucleic acid may comprise a promoter, an intron, an enhancer region, a polyadenylation site, a translation initiation site, 5' or 3' untranslated regions, a reporter gene, a selectable marker or the like. The nucleic acid may comprise single stranded or double stranded DNA or RNA. The nucleic acid may comprise modified bases or a modified backbone. A nucleic acid that is up to about 100 nucleotides in length, is often also referred to as an oligonucleotide. "Nucleotides" as used herein refer to the building blocks of oligonucleotides and polynucleotides, and for the purposes of present disclosure include both naturally occurring and non-naturally occurring nucleotides. In nature, nucleotides, such as DNA and RNA nucleotides comprise a ribose sugar moiety, a nucleobase moiety and one or more phosphate groups (which are absent in nucleosides). A nucleotide without a phosphate group is called a "nucleoside" and is thus a compound comprising a nucleobase moiety and a sugar moiety. As used herein, "nucleobase" means a group of atoms that can be linked to a sugar moiety to create a nucleoside that is capable of incorporation into an oligonucleotide, and wherein the group of atoms is capable of bonding with a complementary naturally occurring nucleobase of another oligonucleotide or nucleic acid. Naturally occurring nucleobases of RNA or DNA comprise the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).

[0044] "Nucleotide sequence", "DNA sequence" or "nucleic acid molecule(s)" as used herein refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. This term refers only to the primary structure of the molecule. Thus, this term includes double- and single-stranded DNA, the (reverse) complement DNA, and RNA. It also includes known types of modifications, for example,methylation, "caps" substitution of one or more of the naturally occurring nucleotides with an analogue. By "nucleic acid construct" it is meant a nucleic acid sequence that has been constructed to comprise one or more functional units not found together in nature. Examples include circular, linear, double-stranded, extrachromosomal DNA molecules (plasmids), cosmids (plasmids containing COS sequences from lambda phage), viral genomes comprising non-native nucleic acid sequences, and the like. "Coding sequence" is a nucleotide sequence, which is transcribed into mRNA and / or translated into a polypeptide when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a translation start codon at the 5'-terminus and a translation stop codon at the 3'-terminus. A coding sequence can include, but is not limited to mRNA, cDNA, recombinant nucleotide sequences or genomic DNA, while introns may be present as well under certain circumstances.

[0045] An "expression cassette" as used herein comprises any nucleic acid construct capable of directing the expression of a gene / coding sequence of interest, which is operably linked to a promoter of the expression cassette. Expression cassettes are generally DNA constructs preferably including (5' to 3' in the direction of transcription): a promoter region, a polynucleotide sequence, homologue, variant or fragment thereof operably linked with the transcription initiation region, and a termination sequence including a stop signal for RNA polymerase and a polyadenylation signal. It is understood that all of these regions should be capable of operating in biological cells, such as prokaryotic or eukaryotic cells, to be transformed. The promoter region comprising the transcription initiation region, which preferably includes the RNA polymerase binding site, and the polyadenylation signal may be native to the biological cell to be transformed or may be derived from an alternative source, where the region is functional in the biological cell. Such cassettes can be constructed into a "vector". The term "vector" or alternatively "vector construct", "expression vector" or "gene transfer vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid molecule to which it has been linked, and includes any vector known to the skilled person, including any suitable type, but not limited to, for instance, plasmid vectors, cosmid vectors, phage vectors, such as lambda phage, viral vectors, such as adenoviral, AAV or baculoviral vectors, or artificial chromosome vectors such as bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC), or Pl artificial chromosomes (PAC). Expression vectors comprise plasmids as well as viral vectors and generally contain a desired coding sequence and appropriate DNA sequences necessary for the expression of the operably linked coding sequence in a particular host organism (e.g., bacteria, yeast,plant, insect, or mammal) or in in vitro expression systems. Cloning vectors are generally used to engineer and amplify a certain desired DNA fragment and may lack functional sequences needed for expression of the desired DNA fragments. The construction of expression vectors for use in transfecting cells is also well known in the art, and thus can be accomplished via standard techniques (see, for example, Sambrook, Fritsch, and Maniatis, in: Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989; Gene Transfer and Expression Protocols, pp.

[0046] 109-128, ed. E. J. Murray, The Humana Press Inc., Clif ton, N .), and the Ambion 1998 Catalog (Ambion, Austin, Tex.).

[0047] The terms "identical" or percent "identity" in the context of two or more nucleic acid or amino acid sequences refer to two or more sequences that are the same or have a specified percentage of nucleotides or amino acid residues respectively that are the same, when compared and aligned (introducing gaps, if necessary) for maximum correspondence, not considering any conservative amino acid substitutions as part of the sequence identity. The percent identity can be measured using sequence comparison software or algorithms or by visual inspection. Various algorithms and software are known in the art that can be used to obtain alignments of nucleotide or amino acid sequences.

[0048] The term "percent sequence identity" or "% sequence identity" or "percent identity" or "% identity" between two polynucleotide or polypeptide sequences refers to the number of identical matched positions shared by the sequences over a comparison window, taking into account additions or deletions (i.e. gaps) that must be introduced for optimal alignment of the two sequences. A matched position is any position where an identical nucleotide or amino acid is presented in both the target and reference sequence. Gaps presented in the target sequence are not counted since gaps are not nucleotides or amino acids. Likewise, gaps presented in the reference sequence are not counted since target sequence nucleotides or amino acids are counted, not nucleotides or amino acids from the reference sequence. One such non-limiting example of a sequence alignment algorithm is the algorithm described in Karlin et al., 1990, Proc. Natl. Acad. Sci., 87:2264-2268, as modified in Karlin et al., 1993, Proc. Natl. Acad. Sci., 90:5873-5877, and incorporated into the NBLAST and XBLAST programs (Altschul et al., 1991, Nucleic Acids Res., 25:3389-3402). In certain aspects, Gapped BLAST can be used as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. BLAST-2, WU-BLAST-2 (Altschul et al., 1996, Methods in Enzymology, 266:460-480), ALIGN, ALIGN-2 (Genentech, South San Francisco, California) or Megalign (DNASTAR) are additional publicly availablesoftware programs that can be used to align sequences. In certain aspects, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (e.g., using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 90 and a length weight of 1, 2, 3, 4, 5, or 6). In certain alternative aspects, the GAP program in the GCG software package, which incorporates the algorithm of Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) can be used to determine the percent identity between two amino acid sequences (e.g., using either a BLOSUM 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5). Alternatively, in certain aspects, the percent identity between nucleotide or amino acid sequences is determined using the algorithm of Myers and Miller (CABIOS, 4:11-17 (1989)). For example, the percent identity can be determined using the ALIGN program (version 2.0) and using a PAM120 with residue table, a gap length penalty of 12 and a gap penalty of 4. One skilled in the art can determine appropriate parameters for maximal alignment by particular alignment software. In certain aspects, the default parameters of the alignment software are used.

[0049] One skilled in the art will appreciate that the generation of a sequence alignment for the calculation of a percent sequence identity is not limited to binary sequencesequence comparisons exclusively driven by primary sequence data. Sequence alignments can be derived from multiple sequence alignments. One suitable program to generate multiple sequence alignments is ClustalW2, available from www.clustal.org. Another suitable program is Clustal Omega, available at www.ebi.ac.uk / jdispatcher / msa / clustalo, as well as MUSCLE, which can be accessed at www.drive5.com / muscle. ClustalW2, Clustal Omega and MUSCLE are alternatively available, e.g., from the EBI (European Bioinformatics Institute).

[0050] In certain aspects, the percentage identity "X" of a first nucleotide sequence to a second nucleotide sequence is calculated as 100 x (Y / Z), where Y is the number of nucleotide residues scored as identical matches in the alignment of the first and second sequences (as aligned by visual inspection or a particular sequence alignment program) and Z is the total number of residues in the second sequence. If the length of a first sequence is longer than the second sequence, the percent identity of the first sequence to the second sequence will be higher than the percent identity of the second sequence to the first sequence. Different regions within a single polynucleotide target sequence that align with a polynucleotide reference sequence can each have their own percent sequence identity. It is noted that the percent sequence identity value is rounded to the nearest tenth. For example, 80.11, 80.12, 80.13, and 80.14 are rounded down to 80.1, while 80.15, 80.16, 80.17, 80.18, and80.19 are rounded up to 80.2. It also is noted that the length value will always be an integer.

[0051] According to the present disclosure, the degree of identity, between a given reference nucleotide sequence and a nucleotide sequence which is a homologue of said given nucleotide sequence will preferably be at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. The degree of identity is given preferably for a nucleic acid region which is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or 100% of the entire length of the reference nucleic acid sequence. For example, if the reference nucleic acid sequence consists of 200 nucleotides, the degree of identity is given preferably for at least 20, at least 40, at least 60, at least 80, at least 100, at least 120, at least 140, at least 160, at least 180, or 200 nucleotides, preferably contiguous nucleotides. In a particular embodiment, the degree / percentage of similarity or identity is given for the entire length of the reference nucleic acid sequence.

[0052] The term "amino acid identity" as used herein refers to the extent that sequences are identical on an amino acid-by-amino acid basis over a window of comparison. Thus, a "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over the window of comparison, determining the number of positions at which the identical amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Vai, Leu, He, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gin, Cys and Met) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. According to the present disclosure, the degree of identity, between a given reference amino acid sequence and an amino acid sequence which is a homologue of said given amino acid sequence will preferably be at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. The degree of identity is given preferably for an amino acid region which is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or 100% of the entire length of the reference amino acid sequence. For example, if the reference amino acid sequence consists of 200 amino acids, the degree of identity is given preferably for at least 20, at least 40, at least 60, at least 80, at least 100, at least 120, at least 140, at least 160, at least 180, or 200 amino acids, preferablycontiguous amino acids. In a particular embodiment, the degree / percentage of similarity or identity is given for the entire length of the reference amino acid sequence.

[0053] "Homologue" or "homologues" of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and / or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived.

[0054] The term "defined by SEQ ID N°: X" or "as depicted in SEQ ID N°: X" as used herein refers to a biological sequence consisting of the sequence of amino acids or nucleotides given in the SEQ ID N°: X. For instance, a protein defined in / by SEQ ID N° : X consists of the amino acid sequence given in SEQ ID N° : X. A further example is an amino acid sequence comprising SEQ ID N°: X, which refers to an amino acid sequence longer than the amino acid sequence given in SEQ ID N°: X but entirely comprising the amino acid sequence given in SEQ ID N°: X (wherein the amino acid sequence given in SEQ ID N°: X can be located N-terminally or C-terminally in the longer amino acid sequence, or can be embedded in the longer amino acid sequence), or to an amino acid sequence consisting of the amino acid sequence given in SEQ ID N°: X.

[0055] A "modulator," as used herein, refers to a molecule (e.g., binding agent, ligand, or antibody) that interacts with a target in a signaling pathway, thereby altering the pathway's activity. Modulation can involve either activation or inhibition of the signaling pathway, depending on the nature of the interaction between the modulator and its target. In the context of the Wnt / beta-catenin signaling pathway, a modulator may either enhance or suppress the stabilization and accumulation of beta-catenin and the subsequent transcriptional activation of Wnt target genes. The modulator may exhibit a range of affinities or specificities for the target, yet still exert an effect on the pathway's activity. In the present context, the term "modulator" encompasses both activators (e.g., agonists) and inhibitors (e.g., antagonists), and includes molecules that selectively influence specific Wnt signaling components, such as the Wnt7 / beta-catenin signaling pathway.

[0056] An "agonist," as used herein, refers to a molecule (e.g., binding agent, ligand, or antibody) that, upon interaction with a receptor or another target in a signaling pathway, activates downstream signaling, thereby potentially initiating a biological response. In the context of the Wnt / beta-catenin signaling pathway, an agonist functions as an activator of this pathway, leading to the stabilization andaccumulation of beta-catenin and subsequent transcriptional activation of Wnt target genes. The agonist may not necessarily exhibit strong binding affinity to the receptor but still effectively promotes pathway activation. In the present context, the term "agonist" may be used interchangeably with the term "activator", preferably of the Wnt / beta-catenin signaling pathway, more preferably of the Wnt7 / beta-catenin signaling pathway.

[0057] Conversely, an "antagonist" is a molecule that interferes with or inhibits the activation of downstream signaling pathways, thereby preventing such biological response. In the context of the Wnt / beta-catenin signaling pathway, an antagonist functions by blocking or inhibiting the pathway, thereby preventing the accumulation of beta-catenin and the transcriptional activation of Wnt target genes. The antagonist does not activate the pathway and may act by preventing the agonist or natural ligand from initiating the signaling cascade. In the present context, the term "antagonist" may be used interchangeably with the term "inhibitor", preferably of the Wnt / beta-catenin signaling pathway, more preferably of the Wnt7 / beta-catenin signaling pathway.

[0058] The terms "treatment" or "treating" or "treat" can be used interchangeably and are defined by a therapeutic intervention that slows, interrupts, arrests, controls, stops, reduces, or reverts the progression or severity of a sign, symptom, disorder, condition, injury, or disease, but does not necessarily involve a total elimination of all disease-related signs, symptoms, conditions, or disorders. Those in need of treatment include those already diagnosed with the disorder as well as those prone or predisposed to contract the disorder or those in whom the disorder is to be prevented. For example, in tumor (e.g. cancer) treatment, a therapeutic agent can directly decrease the pathology of tumor cells, or render the tumor cells more susceptible to treatment by other therapeutic agents or by the subject's own immune system.

[0059] As used herein, the terms "diagnosis", "prognosis" and / or "prediction" comprise diagnosing, prognosing and / or predicting a certain disease and / or disorder, thereby predicting the onset and / or presence of a certain disease and / or disorder, and / or predicting the progress and / or duration of a certain disease and / or disorder, and / or predicting the response of a patient suffering from a certain disease and / or disorder to therapy.

[0060] The term "statistically significantly" different is well known by the person skilled in the art. Statistical significance plays a pivotal role in statistical hypothesis testing. Itis used to determine whether the null hypothesis should be rejected or retained. The null hypothesis is the default assumption that nothing happened or changed. For the null hypothesis to be rejected, an observed result has to be statistically significant, i.e. the observed p-value is less than the pre-specified significance level a. The p-value of a result, p, is the probability of obtaining a result at least as extreme, given that the null hypothesis were true. In one embodiment, a is 0.05. In a more particular embodiment, a is 0.01. In an even more particular embodiment, a is 0.001.

[0061] A peptide, polypeptide or protein can be naturally occurring, e.g., present in or isolated from nature, e.g., produced or expressed natively or endogenously by a cell or tissue and optionally isolated therefrom. A peptide, polypeptide or protein can be recombinant, i.e., produced by recombinant DNA technology, and / or can be, partly or entirely, chemically or biochemically synthesized. Without limitation, a peptide, polypeptide or protein can be produced recombinantly by a suitable host or host cell expression system and optionally isolated therefrom (e.g., a suitable bacterial, yeast, fungal, plant or animal host or host cell expression system), or produced recombinantly by cell-free translation or cell-free transcription and translation, or non-biological peptide, polypeptide or protein synthesis.

[0062] A conservative amino acid substitution is a substitution of one amino acid with another that has similar chemical and physical characteristics. Conservative amino acid substitutions include substitutions within the following groups: valine, alanine and glycine; leucine, valine, and isoleucine; aspartic acid and glutamic acid; asparagine and glutamine; serine, cysteine, and threonine; lysine and arginine; and phenylalanine and tyrosine. The nonpolar hydrophobic amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine and glutamine. The positively charged (i.e., basic) amino acids include arginine, lysine and histidine. The negatively charged (i.e., acidic) amino acids include aspartic acid and glutamic acid. Any substitution of one member of the above-mentioned polar, basic, or acidic groups by another member of the same group can be deemed a conservative substitution. By contrast, a non -conservative substitution is a substitution of one amino acid for another with dissimilar characteristics.

[0063] The term "biologically active" is interchangeable with terms such as "functionally active" or "functional", denoting that the proteins disclosed herein at least partlyretain the biological activity or intended functionality of the respective or corresponding peptide, polypeptide or protein. Reference to the "activity" of a peptide, polypeptide or protein may generally encompass any one or more aspects of the biological activity of the peptide, polypeptide or protein, such as without limitation any one or more aspects of its biochemical activity, enzymatic activity, signaling activity, interaction activity, ligand activity, and / or structural activity, e.g., within a cell, tissue, organ or an organism.

[0064] Preferably, a functionally active protein may retain at least about 20%, e.g., at least about 25%, or at least 30%, or at least about 40%, or at least about 50%, e.g., at least 60%, more preferably at least about 70%, e.g., at least 80%, yet more preferably at least about 85%, still more preferably at least about 90%, and most preferably at least about 95% or even about 100% of the intended biological activity or functionality compared with the corresponding protein.

[0065] In certain embodiments, a functionally active protein may even display higher biological activity or functionality compared with the corresponding peptide, polypeptide or protein, for example may display at least about 100%, or at least about 150%, or at least about 200%, or at least about 300%, or at least about 400%, or at least about 500% of the intended biological activity or functionality compared with the corresponding protein. By means of an example, where the activity of a given protein can be readily measured in an assay with a quantitative output, for example an enzymatic assay or a signaling assay or a binding assay producing a quantifiable signal, a functionally active fragment or variant of the peptide, polypeptide or protein may produce a signal which is at least about 20%, or at least about 25%, or at least 30%, or at least about 40%, or at least about 50%, or at least 60%, more preferably at least about 70%, or at least 80%, or at least about 85%, or at least about 90%, or at least about 95%, or at least about 100%, or at least about 150%, or at least about 200%, or at least about 300%, or at least about 400%, or at least about 500% of the signal produced by the corresponding protein.

[0066] In the present specification, references to any peptides, polypeptides, proteins, or nucleic acids denote the respective peptides, polypeptides, proteins, or nucleic acids as commonly known under the respective designations in the art. More particularly, the references to "Wnt" and in particular to "Wnt7", to "G-protein coupled receptor 124" (GPR124), "Reversion-inducing cysteine-rich protein with Kazal motifs" (RECK), "Frizzled" (FZD), or "lipoprotein receptor-related protein" (LRP) denote therespective peptides, polypeptides, proteins or nucleic acids, as apparent from the context, as commonly known under said designations in the art.

[0067] "Wnt7" in the present context refers to both "Wnt7a" and / or "Wnt7b", which are part of the Wnt-family of proteins. The terms "Wnt7" and "WNT7" are used interchangeably throughout this specification. "Wnt" or "Wingless-related integration site" or "Wingless and Int-1" or "Wingless-Inti" refers to a family of genes and proteins comprising the homologous "Wingless" or "Wg" and "Integration site" or "Int". In a particular embodiment, Wnt is Wnt7a protein with amino acid sequence as depicted in SEQ ID N°: 1. In another particular embodiment, Wnt is Wnt7b protein with amino acid sequence as depicted in SEQ ID N°: 2.

[0068] A "Gprl24 / RECK / FZD / LRP receptor complex", "Gprl24 / RECK / FZD / LRP co-receptor complex", "Gprl24 / RECK / FZD / LRP complex" or "Gprl24 / RECK / FZD / LRP protein complex" broadly denotes a protein complex, particularly a membrane-associated protein complex, more particularly a plasma membrane-associated protein complex comprising at least one Gprl24 protein, at least one RECK protein, at least one FZD protein and at least one LRP protein. A GPR124 / RECK / FZD / LRP receptor complex, when located at the plasma membrane of a cell, is capable of activating Wnt / 0-catenin signaling in said cell in response to extracellularly provided Wnt7 ligand.

[0069] A "FZD / LRP receptor complex", "FZD / LRP co-receptor complex", "FZD / LRP complex" or "FZD / LRP protein complex" broadly denotes a protein complex, particularly a membrane-associated protein complex, more particularly a plasma membrane associated protein complex comprising at least one FZD protein and at least one LRP protein. A FZD / LRP receptor complex, when located at the plasma membrane of a cell, is capable of activating Wnt / 0-catenin signaling in said cell in response to extracellularly provided Wnt ligand, such as, but not limited to Wnt7 ligand.

[0070] "Wnt signaling" and "Wnt / 0-catenin signaling" are used interchangeably herein.

[0071] "Frizzled", "Fz", "FZ", "Fzd" or "FZD" as used herein denotes a family of at least ten atypical G protein-coupled receptors (GPCR) that serve as receptors in the Wnt signaling pathway and other signaling pathways. When activated, Fzd leads to activation of Dishevelled in the cytosol. In certain particularly preferred embodiments, the terms denote Fzdl, Fzd4, Fzd5 and / or Fzd8.

[0072] In a particular embodiment, Fzd is Fzdl, wherein Fzdl is the protein with amino acid sequence as depicted in SEQ ID N°: 9."LRP" or "lipoprotein receptor-related protein" encompasses any and all lipoprotein receptor-related proteins, also known in the art as low-density lipoprotein receptor-related proteins or prolow-density lipoprotein receptor- related proteins. In certain particularly preferred embodiments, the terms denote LR.P5, LR.P6, or LR.P5 and LR.P6 (LRP5 / 6). In a particular embodiment, LRP is LRP5 with amino acid sequence as depicted in SEQ ID N°: 3. In another particular embodiment, LRP is LRP6 with amino acid sequence as depicted in SEQ ID N°: 4 or 5. As both LRP5 and LRP6 may exist as multiple isoforms, a person skilled in the art will understand that these isoforms are also understood as comprised herein.

[0073] The interaction between Wnt ligands, Fzd receptors, and LRP is essential for the activation of the canonical Wnt pathway, which has wide-ranging effects on cell proliferation, differentiation, and tissue development. This binding event triggers a series of intracellular signaling events that ultimately lead to the stabilization and nuclear translocation of a protein called 0-catenin. In the nucleus, 0-catenin functions as a transcriptional co-activator, regulating the expression of target genes involved in various cellular processes.

[0074] "Reck", or "RECK" is a GPI-anchored membrane protein that stands for "Reversioninducing Cysteine-rich Protein with Kazal Motifs". In a particular embodiment, Reck is the protein with amino acid sequence as depicted in SEQ ID N°: 6.

[0075] "Gprl24" or "GPR124" is an orphan GPCR expressed in endothelial cells. Gprl24 is also named ADGRA2 or adhesion G protein-coupled receptor A2. In a particular embodiment, Gprl24 is the protein with amino acid sequence as depicted in SEQ ID N°: 7.

[0076] The terms "bind", "interact", "specifically bind" or "specifically interact" as used throughout this specification mean that an agent binds to or influences one or more desired molecules or analytes substantially to the exclusion of other molecules which are random or unrelated, and optionally substantially to the exclusion of other molecules that are structurally related. The terms do not necessarily require that an agent binds exclusively to its intended target(s). For example, an agent may be said to specifically bind to target(s) of interest if its affinity for such intended target(s) under the conditions of binding is at least about 2-fold greater, preferably at least about 5-fold greater, more preferably at least about 10-fold greater, yet more preferably at least about 25-fold greater, still more preferably at least about 50-fold greater, and even more preferably at least about 100-fold or more greater, such as,e.g., at least about 1000-fold or more greater, at least about lxl04-fold or more greater, or at least about lxl05-fold or more greater, than its affinity for a nontarget molecule.

[0077] The term "gene therapy" and / or "RIMA therapy" as used herein refers to the introduction of an exogenous polynucleotide into a host cell for therapeutic or prophylactic purposes, irrespective of the method used for the introduction. Such methods include a variety of well-known techniques such as vector-mediated gene transfer (by, e.g., viral infection / transfection, or various other protein-based or lipid-based gene delivery complexes) as described elsewhere herein. The introduced polynucleotide may be stably or transiently maintained in the host cell. Stable maintenance typically requires that the introduced polynucleotide either contains an origin of replication compatible with the host cell or integrates into a replicon of the host cell such as an extrachromosomal replicon (e.g., a plasmid) or a nuclear or mitochondrial chromosome. A number of vectors are known to be capable of mediating transfer of genes to mammalian cells, as is known in the art. The RNA molecule may be any type of RNA molecule. For instance, the RNA molecule may be cytoplasmic RNA, a nuclear RNA, an mRNA, antisense RNA or a noncoding RNA, preferably an mRNA.

[0078] The terms "host cell" and "host organism" may suitably refer to cells or organisms encompassing both prokaryotes, such as bacteria, and eukaryotes, such as yeast, fungi, protozoan, plants and animals. Contemplated as host cells are inter alia unicellular organisms, such as bacteria (e.g., E. coli, Salmonella typhimurium, Serratia marcescens, or Bacillus subtilis), yeast (e.g., Saccharomyces cerevisiae or Pichia pastoris), (cultured) plant cells (e.g., from Arabidopsis thaliana or Nicotiana tabacum) and (cultured) animal cells (e.g., vertebrate animal cells, mammalian cells, primate cells, human cells or insect cells). Contemplated as host organisms are inter alia multi-cellular organisms, such as plants and animals, preferably animals, more preferably warm-blooded animals, even more preferably vertebrate animals, still more preferably mammals, yet more preferably primates; particularly contemplated are such animals and animal categories which are non-human.

[0079] Except when noted, the terms "subject" or "patient" can be used interchangeably and refer to animals, preferably warm-blooded animals, more preferably vertebrates, even more preferably mammals, still more preferably primates, and specifically includes human patients and non-human mammals and primates. Preferred subjects are human subjects. The terms "subject" or "patient" includesubjects in need of treatment, more particularly subjects that would benefit from treatment of a given condition, particularly a neurological disorder, neurovascular disorder or a central nervous system (CNS) disorder comprising neurovascular dysfunction; or to treat blood-retina barrier (BRB) integrity, or the treatment of an ophthalmological disease or disorder, such as a retinal vascular disorder or retinal degenerative disorder. Such subjects may include, without limitation, those that have been diagnosed with said condition, those prone to develop said condition and / or those in who said condition is to be prevented.

[0080] The term "therapeutically active / effective amount" as used herein, refers to an amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a subject that is being sought by a surgeon, researcher, veterinarian, medical doctor or other clinician, which may include inter alia alleviation of the symptoms of the disease or condition being treated. The term "prophylactically active / effective amount" refers to an amount of an active compound or pharmaceutical agent that inhibits or delays in a subject the onset of a disorder as being sought by a researcher, veterinarian, medical doctor or other clinician. In present context, the "therapeutically active amount" is used to refer to both the "therapeutically active amount" and "prophylactically active amount" as described above, unless the distinction is clear from the context. Methods are known in the art for determining therapeutically and / or prophylactically effective doses of a compound, a protein, a nucleic acid encoding the compound / protein, a nucleic acid expression cassette, or a pharmaceutical composition, as taught herein. The term "therapeutically effective dose" as used herein refers to an amount of a compound, a protein, a nucleic acid encoding the compound / protein, a nucleic acid expression cassette, or a pharmaceutical composition, as taught herein, that when administered brings about a positive therapeutic response with respect to treatment of a patient having a specific disease or disorder.

[0081] Detailed description

[0082] The inventors demonstrate herein the design of modulators such as agonists capable of activating Wnt7 signaling selectively in cells expressing RECK and / or GPR124; for instance cells expressing GPR124, or cells expressing RECK, or cells expressing GPR124 and RECK i.e. for instance in cerebral or retinal endothelial cells. It is further disclosed that such modulators are useful as therapeutics, such as particularly for the treatment of neurological disorders, neurovascular disorders or central nervous system (CNS) disorders comprising neurovascular dysfunction, or an ophthalmological disease or disorder in particular a retinal vascular disorder orretinal degenerative disorder. Hence, the invention allows providing inter alia agonists capable of stimulating Wnt / 0-catenin signaling in cerebral or retinal endothelial cells (also referred to as "on-target activity") with only low crossreactivity with other Fzd pathways (i.e. "unwanted" or "off-target activity"), and useful as therapeutics, particularly for neurological disorders, neurovascular disorders or central nervous system (CNS) disorders comprising neurovascular dysfunction, and / or for ophthalmological diseases or disorders such as retinopathies. In particular, said modulators are preferably useful as therapeutics for use in the treatment of disorders linked to the blood-brain barrier integrity or for use in the treatment of the blood-brain barrier integrity, and / or for use in the treatment of disorders linked to the blood-retina barrier integrity or for use in the treatment of the blood-retina barrier integrity.

[0083] The inventors surprisingly discovered that the direct recruitment or binding of the antibody or antibody fragment, i.e. the modulator or agonist, to Fzd and either Gprl24 or RECK; or Fzd, Gprl24 and Reck, is sufficient for stimulating Wnt7 signaling specifically through the Gprl24 / RECK / FZD / LRP receptor complex.

[0084] In the following passages, different aspects or embodiments of the invention are defined in more detail. Each aspect or embodiment so defined may be combined with any other aspect(s) or embodiment(s) unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

[0085] Accordingly, in a first aspect, the invention provides a modulator of the Wnt7 signaling pathway. Said modulator preferably is a, preferably soluble, antibody or antibody fragment. Said modulator preferably is an agonist of the Wnt7 signaling pathway. Accordingly, preferably, the invention provides a soluble antibody or antibody fragment, wherein the antibody or antibody fragment is a modulator of Wnt7 signaling pathway, preferably an agonist of the Wnt7 pathway. The antibody or antibody fragment of the present invention comprises one or more Fzd binding domains, and one or more Gprl24 or Reck binding domains.

[0086] In one embodiment, said antibody or antibody fragment is a modulator, preferably an agonist, preferably a selective or specific agonist, of the Gprl24- and Reck-dependent Wnt / p-catenin pathway. The term "selective agonist of the Gprl24- and Reck-dependent Wnt / p-catenin pathway" or "specific agonist of the Gprl24- andReck-dependent Wnt / p-catenin pathway" is to be understood as an agonist of said pathway that is able to activate said pathway with high selectivity and specificity (on-target activity), with no, no statistically significantly or only minimal activation of other Wnt signaling pathways (off-target activity). In embodiments, the agonist disclosed herein is an agonist of Gprl24- and Reck-dependent Wnt / 0-catenin pathway that shows less than 20% off-target activity, more particularly less than 15%, 10%, 5% or 1% off-target activity, as measured in cell culture Super TOP-Flash assays (as known it the art and discussed below).

[0087] In certain embodiments, one or more and preferably all of Wnt7, GPR124, RECK, FZD and LRP as employed herein is or are of animal origin, preferably warm-blooded animal origin, more preferably vertebrate origin, yet more preferably mammalian origin, including human origin and non-human mammalian origin, still more preferably human origin.

[0088] A skilled person can appreciate that any sequences represented in sequence databases or in the present specification may be precursors of the respective peptides, polypeptides, proteins, or nucleic acids referred to in the present text, and may include parts that are processed away from mature molecules; or may represent sequences that are of mature molecules while also the precursor sequences are considered part of the present concept. The terms encompass the peptides, polypeptides, proteins, or nucleic acids when forming a part of a living organism, organ, tissue, or cell, when forming a part of a biological sample, as well as when at least partly isolated from such sources. The terms also encompass the peptides, polypeptides, proteins, or nucleic acids when produced by recombinant or synthetic means.

[0089] Unless otherwise apparent from the context, reference herein to any peptide, polypeptide, protein, or nucleic acid also encompasses modified forms of said peptide, polypeptide, protein, or nucleic acid, such as forms bearing post-expression modifications including, for example, phosphorylation, glycosylation, palmitoylation, lipidation, methylation, cysteinylation, sulphonation, glutathionylation, acetylation, ubiquitination, oxidation of methionine to methionine sulphoxide or methionine sulphone, signal peptide removal, N-terminal Met removal, conversion of proenzymes or pre-hormones into active forms, and the like. A broader definition is given above.The antibodies and antibody fragments as disclosed herein may have any of a variety of different structural formats or configurations. Said antibodies or antibody fragments may comprise one or more heavy chains and / or one or more light chains, or they may be single-domain antibodies such as VHH domains. In particular embodiments, said antibodies or antibody fragments may comprise a single polypeptide, or they may comprise two or more, three or more, or four or more polypeptides. In certain embodiments, said antibodies or antibody fragments may comprise one, two, three, or four polypeptides, e.g., linked or bound to each other or fused to each other.

[0090] When the antibody or antibody fragment comprises a single polypeptide, it may be a fusion protein comprising one or more antigen-binding regions, such as a heavy chain variable domain (VH) or a light chain variable domain (VL), and optionally other binding domains or functional regions. The binding regions may be directly fused or they may be connected via a linker, e.g., a polypeptide or chemical linker, including but not limited to any of those disclosed herein.

[0091] When the antibody or antibody fragment comprises two or more polypeptides, the polypeptides may be linked via covalent bonds, such as, e.g., disulfide bonds, and / or noncovalent interactions. For example, heavy chains of human immunoglobulin IgG interact at the level of their CH3 domains directly, whereas, at the level of their CH2 domains, they interact via the carbohydrate attached to the asparagine (Asn) N84.4 in the DE turn.

[0092] In an embodiment, the antibodies or antibody fragments as described herein may be engineered.

[0093] In another or further embodiment, the antibodies or antibody fragments as described herein may be water soluble. By "water soluble", or short "soluble" it is meant a composition that is soluble in aqueous buffers in the absence of detergent, usually soluble at a concentration that provides a biologically effective dose of the antibody or antibody fragment. The antibodies or antibody fragments disclosed herein typically form a substantially homogeneous aqueous solution at concentrations of at least 2.5 pM, and higher, e.g., at least 2.5 pM, 3 pM, 4 pM, 5 pM, 10 pM, 15 pM, 20 pM, 25 pM, 40 pM, or 50 pM, and in some embodiments at least 60 pM, 70 pM, 80 pM, or 90 pM, sometimes as much as 100 pM, 120 pM, or 150 pM. In other words, the antibodies or antibody fragments disclosed herein typically form a substantially homogeneous aqueous solution at concentrations of about 0.5 mg / ml, about 1.0mg / ml, about 1.5 mg / ml, about 2.0 mg / ml, about 2.5 mg / ml, about 5.0 mg / ml, about 10.0 mg / ml, about 15.0 mg / ml, about 20.0 mg / ml, about 25.0 mg / ml, about 30.0 mg / ml, or about 35.0 mg / ml or more.

[0094] The antibodies or antibody fragments as described herein may be engineered to facilitate binding between two polypeptides. For example, knobs-into-holes amino acid modifications may be introduced into two different polypeptides to facilitate their binding. Knobs-into-holes amino acid (AA) changes is a rational design strategy developed in antibody engineering, used for heterodimerization of the heavy chains, in the production of bispecific IgG antibodies. AA changes are engineered in order to create a knob on the CH3 domain of the heavy chains from a first antibody and a hole on the CH3 domain of the heavy chains of a second antibody. The knob may be represented by a tyrosine (Y) that belongs to the 'very large' IMGT volume class of AA, whereas the hole may be represented by a threonine (T) that belongs to the 'small' IMGT volume class. Other means of introducing modifications into antibodies or antibody fragments to facilitate their binding are known and available in the art. For example, specific amino acids may be introduced and used for cross-linking, such as cysteine to form an intermolecular disulfide bond.

[0095] In particular embodiments, the antibodies or antibody fragments comprise one or more binding regions derived from an antibody, such as a full-length immunoglobulin, an scFv, a Fab fragment, or a single-domain antibody (VHH, VNAR, or other sdAbs), e.g., antibody heavy chains or antibody light chains or fragments thereof.

[0096] In an embodiment, said antibody or antibody fragment is capable of binding to Fzd, and further to Gprl24 and / or to Reck. Said antibody or antibody fragment is thus preferably capable of binding to Fzd and Gprl24, to Fzd and Reck, or to Fzd, Gprl24, and Reck.

[0097] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains and one or more Reck binding domains, or one or more Fzd binding domains and one or more Gprl24 binding domains, or one or more Fzd binding domains and one or more Reck binding domains and one or more Gprl24 binding domains.

[0098] Optionally, the antibody or antibody fragment has further binding domains.Preferably, the antibody or antibody fragment has no further binding domains over said Fzd and Gprl24, Fzd and Reck, or Fzd, Gprl24, and Reck binding sites.

[0099] In an embodiment, the antibody or antibody fragment as disclosed herein is capable of simultaneously binding to the Fzd protein, and to GPR124 and / or RECK proteins. In particular the antibody or antibody fragment as disclosed herein is capable of simultaneously binding to the Fzd protein and Gprl24 protein, to the Fzd protein and Reck protein, or to the Fzd protein, Gprl24 protein and Reck protein.

[0100] In an embodiment, the antibody or antibody fragment as disclosed herein is capable of binding to epitopes located in the extracellular domains of Fzd, GPR124 and / or RECK.

[0101] In an embodiment, the antibody or antibody fragment, which is a modulator of the Wnt7 signaling pathway, comprises one or more Fzd binding domains, and further one or more Gprl24 and / or Reck binding domains. In a further embodiment, said antibody or antibody fragment comprises one, two, three, four, or five Fzd binding domains; and further one, two, three, four, or five Gprl24 binding domains, and / or one, two, three, four, or five Reck binding domains.

[0102] In a preferred embodiment, said antibody or antibody fragment comprises one or two Fzd binding domains; one or two Gprl24 binding domains, and / or one or two Reck binding domains.

[0103] In an embodiment, at least two of the binding domains of the antibody or antibody fragment are fused in tandem, optionally separated by a linker sequence.

[0104] Said linkers may be any linker known in the art, or described in this text. As a nonlimiting example, said linker is a 5-mer linker such as GSGGS (SEQ ID N°: 172).

[0105] In an embodiment, said antibody or antibody fragment comprises at least one region that specifically binds to Fzd epitope, and at least one region that specifically binds to a Gprl24 epitope or to a Reck epitope.

[0106] In an embodiment, said antibody or antibody fragment is bispecific.

[0107] In an embodiment, where the antibody or antibody fragment is bispecific, they may be assembled by fusing a scFv to an IgG, using the DVD-Ig format, or any format asdescribed below. A person skilled in the art will understand that further formats are possible as well. Non-limiting examples are provided below. In the below examples, it is considered clear that specificity for the Fzd epitope can be considered specificity for the first or second epitope, while specificity for the Reck or Gprl24 epitope can be considered specificity for the second or first epitope respectively.

[0108] For example, in an embodiment, a bispecific antibody may be generated by fusing an scFv to an IgG scaffold. The scFv, which consists of a variable heavy (VH) and variable light (VL) domain connected by a linker, may be fused to the N- or C-termini of the heavy chain or the light chain of the IgG using linkers of various lengths. This configuration enables binding to one epitope via the IgG Fab region, while the scFv provides specificity to a second epitope. In the present example, this configuration enables binding to a Fzd epitope via the IgG Fab region, while the scFv provides specificity to a second epitope, such as a Gprl24 epitope or a Reck epitope; or that this configuration enables binding to a Gprl24 epitope or a Reck epitope via the IgG Fab region, while the scFv provides specificity to a second epitope, such as a Fzd epitope.

[0109] In another embodiment, the bispecific antibody or antibody fragment is generated using a Dual Variable Domain Immunoglobulin (DVD-Ig) format. In this format, each Fab arm comprises two VH and two VL domains arranged in tandem, such that the outermost VH / VL pair provides specificity for a first epitope., while the innermost VH / VL pair provides specificity for a second epitope. This design allows for the simultaneous targeting of two different antigens within a single Fab arm, while maintaining the IgG-like structure.

[0110] Further embodiments include alternative bispecific formats, such as the Knobs-into-Holes (KiH) format, wherein Fc heterodimerization is promoted by introducing steric mutations in the CH3 domain, thereby enabling the pairing of distinct heavy chains. Similarly, the CrossMab format may be employed, wherein domain swapping between the CHI and CL regions ensures correct heavy-light chain pairing, reducing the risk of mispairing.

[0111] In an embodiment, the bispecific antibody or antibody fragment may be assembled using a tandem scFv format, in which two scFv domains specific for distinct epitopes are linked in series via a flexible peptide linker, forming a compact, flexible bispecific molecule. Alternatively, a Fab-scFv fusion format may be used, wherein one Fab armretains its natural antigen-binding function while the second Fab is replaced or extended with an scFv targeting a second epitope.

[0112] A person skilled in the art will appreciate that these formats, along with additional known bispecific antibody architectures, may be employed to achieve the desired specificity and functionality while maintaining favorable biophysical and pharmacokinetic properties.

[0113] In an embodiment, where the antibody or antibody fragment is bispecific, the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains and one or more Gprl24 binding domains. Preferably, said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains and one or more Gprl24 binding domains.

[0114] In another embodiment, where the antibody or antibody fragment is bispecific, the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains and one or more Reck binding domains. Preferably, said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains and one or more Reck binding domains.

[0115] In an embodiment, said antibody or antibody fragment is a bispecific antibody with two binding domains, said binding domains including one Fzd binding domain, and one Gprl24 binding domain or Reck binding domain.

[0116] In an embodiment, said antibody or antibody fragment is trispecific.

[0117] In embodiment, where said antibody or antibody fragment is trispecific, said antibody or antibody fragment comprises at least one region that specifically binds to Fzd epitope, at least one region that specifically binds to a Gprl24 epitope, and at least one region that specifically binds to a Reck epitope.

[0118] In embodiments where more than two epitopes are targeted, the antibody or antibody fragment may adopt a trispecific or multispecific format, such as a TriTAC (trispecific T-cell activating construct) ora heavy-chain-only antibody (HCAb) fusion, designed to engage additional molecular targets.A person skilled in the art will understand that further formats are possible as well. Non-limiting examples are provided below. In these examples, it is understood that specificity for the Fzd, Gprl24, or Reck epitope can independently be considered specificity for the first, second, or third epitope, respectively. In other words, the order of epitope specificity is not limited to the configurations described, and each epitope can occupy any of the three binding positions depending on the desired functional arrangement.

[0119] Thus, in an embodiment, the antibody or antibody fragment is trispecific, capable of simultaneously binding to Fzd, Gprl24, and Reck epitopes. Such trispecific formats allow for coordinated engagement of all three targets, potentially enhancing therapeutic efficacy by orchestrating complex signaling cascades.

[0120] In one embodiment, the trispecific antibody is configured as an IgG scaffold engineered with two distinct binding regions on each Fab arm, enabling simultaneous binding to for instance Fzd and Gprl24, while an additional binding moiety specific for for instance Reck is fused to the IgG Fc C-terminal region. Another design leverages a scFv-IgG fusion, where the scFv targeting Fzd is fused to the N-terminus of one heavy chain, the scFv targeting Gprl24 is fused to the N-terminus of the other heavy chain, and an additional binding domain for Reck is linked to the Fc C-terminal region. Such configurations facilitate spatial arrangement and avidity effects, enhancing multi-target engagement.

[0121] In another embodiment, the trispecific antibody is assembled using a Dual Variable Domain Immunoglobulin (DVD-Ig) format. In this configuration, one Fab arm comprises two VH and two VL domains arranged in tandem, with the outermost VH / VL pair binding Fzd and the innermost VH / VL pair binding Gprl24. The opposing Fab arm is engineered to specifically bind Reck, enabling simultaneous engagement of all three targets.

[0122] In an embodiment, the trispecific antibody is assembled using a tandem scFv format, wherein three scFv domains specific for Fzd, Gprl24, and Reck are linked in series via flexible peptide linkers. This format yields a compact, flexible trispecific molecule, allowing simultaneous engagement while maintaining favorable pharmacokinetic properties.In yet another embodiment, the trispecific antibody may be configured as a TriTAC (trispecific T-cell activating construct), designed to engage Fzd, Gprl24, and Reck.

[0123] A person skilled in the art will appreciate that these trispecific formats, as well as additional known architectures, may be employed to achieve the desired specificity and functionality while maintaining favorable biophysical and pharmacokinetic properties.

[0124] In an embodiment, said antibody or antibody fragment is trispecific and / or trivalent.

[0125] In an embodiment, said antibody or antibody fragment comprises one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains.

[0126] In an embodiment, where said antibody or antibody fragment is trispecific, it comprises:

[0127] i) one region that specifically binds to a set of one or more Fzd epitopes; ii) one region that specifically binds to a set of one or more Gprl24 epitopes;

[0128] and

[0129] iii) one region that specifically binds to a set of one or more Reck epitopes.

[0130] In an embodiment, where said antibody or antibody fragment is trispecific, the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains.

[0131] In an embodiment, said antibody or antibody fragment is a trispecific antibody with three binding domains, said binding domains including one Fzd binding domain, one Gprl24 binding domain, and one Reck binding domain.

[0132] In an embodiment, the one or more Gprl24 binding domains bind an amino acid sequence having at least 75% sequence identity to a fragment of the Gprl24 proteins as depicted in SEQ ID N°: 7, or at least 80%, or at least 85%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 98.1%, or at least 98.2%, or at least 98.3%, or at least 98.4%, or at least 98.5%, or at least 98.6%, or at least 98.7%, or at least 98.8%, or at least 98.9%, or at least 99%, orat least 99.1%, or at least 99.2%, or at least 99.3%, or at least 99.4%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%.

[0133] In an embodiment, the one or more Reck binding domains bind an amino acid sequence having at least 75% sequence identity to a fragment of the Reck protein as depicted in SEQ ID N°: 6, or at least 80%, or at least 85%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 98.1%, or at least 98.2%, or at least 98.3%, or at least 98.4%, or at least 98.5%, or at least 98.6%, or at least 98.7%, or at least 98.8%, or at least 98.9%, or at least 99%, or at least 99.1%, or at least 99.2%, or at least 99.3%, or at least 99.4%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%.

[0134] In an embodiment, the one or more Fzd binding domains bind an amino acid sequence having at least 75% sequence identity to a fragment of the Fzd protein as depicted in SEQ ID N°: 9, or at least 80%, or at least 85%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 98.1%, or at least 98.2%, or at least 98.3%, or at least 98.4%, or at least 98.5%, or at least 98.6%, or at least 98.7%, or at least 98.8%, or at least 98.9%, or at least 99%, or at least 99.1%, or at least 99.2%, or at least 99.3%, or at least 99.4%, or at least 99.5%, or at least 99.6%, or at least 99.7%, or at least 99.8%, or at least 99.9%.

[0135] In an embodiment, in at least one of the one or more Gprl24 binding domains and / or the one or more Reck binding domains and / or one or more Fzd binding domains are derived from an antibody, an antibody fragment, an immunoglobulin single variable domain (ISVD) or VHH.

[0136] In an embodiment, said antibody or antibody fragment is not capable of or has an impaired capacity to bind to LRP such as LRP5 or LRP6, or any of the further known LRP proteins, and / or does not comprise functional LRP binding domains, or does not comprise LRP binding domains. In another or further embodiment, said antibody or antibody fragment is not able to or has an impaired capacity to activate LRP. In the present context, LRP is meant to refer to LRP5 or LRP6, or any of the further known LRP proteins involved in the Wnt / 0-catenin pathway.In an embodiment, said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains, and said one or more Gprl24 binding domains and / or one or more Reck binding domains.

[0137] In an embodiment, the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains, and further one or more Gprl24 binding domains and / or one or more Reck binding domains.

[0138] In an embodiments, the antibody or antibody fragment as disclosed herein cannot stimulate the Wnt7 signaling pathway in cells which are GPR124 / RECK negative. In further embodiments, GPR124 / RECK negative means having no or low expression of GPR124 and / or RECK proteins, no or low expression of GPR124 and / or RECK in a cell, no or low presence of GPR124 and / or RECK proteins in a cell or no or low presence of GPR124 and / or RECK proteins at a cell surface. In embodiments, GPR124 / RECK positive refers to the expression of GPR124 and / or RECK in a cell, presence of GPR124 and / or RECK proteins in a cell and / or presence of GPR124 and / or RECK proteins at a cell surface.

[0139] In other or further embodiments, FZD / LRP positive can mean the expression of FZD and / or LRP in a cell, presence of FZD and / or LRP protein in a cell, and / or presence of FZD and / or LRP proteins at a cell surface.

[0140] In particular embodiments, cells expressing GPR124, RECK, FZD and LRP proteins, more particularly LRP5 or LRP6, at its plasma membrane, are cells naturally expressing all GPR124, RECK, FZD and LRP proteins at the cell surface, such as a cerebral / retinal endothelial cell.

[0141] In some embodiments "the presence of RECK and GPR124" may refer to the presence of the RECK and GPR124 proteins at or close to the cell membrane, preferably in close proximity with the Fzd and LRP proteins, more particularly LRP5 or LRP6. The close mutual proximity can facilitate the formation of the GPR124 / RECK / Fzd / LRP receptor complex under conditions conducive thereto, such as when the antibody or antibody fragment as taught herein is externally supplied to a cell. In other or further embodiments, when used in gene or RNA therapy, the antibody or antibody fragment as taught herein could be expressed from the target cell population. On the other hand, the phrase "the absence of RECK and / or GPR124" may refer to the lack or non-occurrence of the RECK and / or GPR124 proteins at the cell membrane. The absence of RECK and / or GPR124 at or close to the cellmembrane may occur when a cell does not express, translate, or correctly translocate RECK and / or GPR124. The absence of RECK and / or GPR124 need not denote the complete absence of the RECK and / or GPR124 protein at the cell membrane, but may for example refer to an amount of RECK and / or GPR124 protein which is not detectable by, or falls below the sensitivity range of, conventional protein detection or quantification assays known by the person skilled in the art, such as for example immunoblotting, immunocytochemistry or immunofluorescence.

[0142] In other or further embodiments, said antibody or antibody fragment herein disclosed is capable of activating the GPR124 / RECK / Fzd / LRP-mediated Wnt7 signaling, wherein said antibody or antibody fragment does not or not significantly activate Fzd / LRP-mediated Wnt signaling in the absence of RECK and / or GPR124.

[0143] In particular embodiments, the capability of the antibody or antibody fragment herein disclosed to activate GPR124 / RECK / Fzd / LRP-mediated Wnt7 signaling, but not or not significantly activate Fzd / LRP-mediated Wnt signaling in the absence of RECK and / or GPR124, denotes the capability of the antibody or antibody fragment to activate Wnt7 signaling in cells positive for GPR124, RECK, FZD and LRP, more particularly LRP5 or LRP6, but not or not significantly in cells positive for FZD and LRP and negative for GPR124 and / or RECK, wherein the cells positive for GPR124, RECK, FZD and LRP, more particularly LRP5 or LRP6, and the cells positive for FZD and LRP and negative for GPR124 and / or RECK are otherwise substantially identical.

[0144] In particular embodiments, the capability of the antibody or antibody fragment to activate GPR124 / RECK / Fzd / LRP-mediated Wnt7 signaling, but not or not significantly activate the canonical Fzd / LRP-mediated Wnt signaling in the absence of RECK and / or GPR124, denotes the capability of said antibody or antibody fragment to activate the atypical Wnt signaling in the presence of RECK and GPR124, but not or not significantly in the absence of RECK and / or GPR124.

[0145] The capability of activating Wnt7 signaling refers to the ability of the antibody or antibody fragment as disclosed herein to mimic, reproduce or approximate the signal transduction effect and / or activity of a natural Wnt7 ligand binding to a Gprl24 / RECK / FZD / LRP complex.

[0146] Activation of Wnt7 signaling may be suitably determined and / or quantitated by measuring the expression of one or more Wnt7 target genes, TCF reporter gene expression, beta-catenin stabilization, LRP phosphorylation, and / or translocation ofAxin from cytoplasm to cell membrane as known in the art. For instance, activation of Wnt signaling may be suitably determined and / or quantitated by measuring the expression of TCF gene (e.g., by RT-PCR or any other transcript detection method), a primary output of Wnt signaling. For example, a TCF reporter assay (also known as TOP / FOP or TOP flash) may be used to assess changes in the transcription of TCF / LEF controlled genes. The TCF reporter assay may be a luciferase reporter assay. Further for example, activation of Wnt signaling may be suitably determined and / or quantitated by measuring the expression of c-myc, n-myc, LEF1, or c-jun. Alternatively, activation of Wnt signaling may be determined by measuring the location, level and / or phosphorylation status of 0-catenin. A non-limiting example of such an assay is the "0-Catenin Redistribution Assay” (Thermo Scientific) which provides recombinant U20S cells stably expressing human p-catenin fused to the C-terminus of enhanced green fluorescent protein (EGFP). The assay allows visualization and monitoring of the translocation of a GFP-p-catenin fusion protein from the membrane to the nucleus. Another way of determining activation of Wnt signaling is the visualization of Axin translocation, for example with a GFP-Axin fusion protein.

[0147] In particular embodiments, the antibody or antibody fragment as disclosed herein may be considered capable of activating the central nervous system (CNS) Wnt7 signaling, the retinal Wnt7 signaling, or the Gprl24 / RECK / FZD / LRP5 or LRP6 dependent Wnt7 signaling, if the antibody or antibody fragment enhances Wnt / 0-catenin signaling at least 10-fold more, at least 20-fold more, at least 30-fold more, at least 40-fold more, at least 50-fold more, at least 100-fold more, at least 250-fold more, at least 500-fold more, at least 750-fold more, at least 1000-fold more, at least lxl04-fold more, or at least lxl05-fold more compared to Wnt / 0-catenin signaling baseline or background induced by a neutral substance or negative control, for example as measured in an assay as described elsewhere herein.

[0148] In particular embodiments, the antibody or antibody fragment as disclosed herein may be considered to not or not significantly activate (canonical) Wnt signaling if the compound enhances Wnt / 0-catenin signaling less than 10-fold more, such as particularly at most 5-fold more or at most 2.5-fold more, or if the protein does not enhance or even reduces (e.g., 2-fold less or 5-fold less or 10-fold less) Wnt / 0-catenin signaling compared to Wnt / 0-catenin signaling baseline or background induced by a neutral substance or negative control, for example as measured in an assay as described elsewhere herein.In particular embodiments, the antibody or antibody fragment as disclosed herein may be considered to activate the GPR124 / RECK / Fzd / LRP5 or LRP6-mediated Wnt7 signaling (denoted "on-target activity"), but not activate Fzd / LRP-mediated Wnt signaling in the absence of RECK and / or GPR124 (denoted "off-target activity"), if the on-target signaling activity is induced at least 30% and off-target activity is induced less than 20% by said antibody or antibody fragment. In embodiments, said antibody or antibody fragment can activate GPR124 / RECK / Fzd / LRP5 or LRP6-mediated Wnt7 signaling more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 99% and all ranges and subranges therein, preferably more than 70% compared to a control situation, for example in the absence of said antibody or antibody fragment.

[0149] In embodiments, said antibody or antibody fragment can activate Fzd / LRP-mediated Wnt in the absence of RECK and / or GPR124 less than 20%, less than 18%, less than 16%, less than 14%, less than 12%, less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1% and all ranges and subranges therein, preferable less than 10% compared to a control situation, for example in the presence of Wnt7. Before comparing the GPR124 / RECK / Fzd / LRP-mediated Wnt signaling activity (denoted "on-target activity") and the Fzd / LRP-mediated Wnt signaling activity in the absence of RECK and / or GPR124 (denoted "off-target activity" in this paragraph), on-target activity and off-target activity induced by the antibody or antibody fragment may be normalized to on-target activity induced by wild-type Wnt7a or b, the latter for example set to represent 100% activity.

[0150] In one embodiment, the antibody or antibody fragment of the present invention acts by Wnt7-specific RECK / GPR124 / Fzd / LRP5 or LRP6-mediated signaling, in which said antibody or antibody fragment binds specifically to GPR124 and / or RECK in an LRP-independent manner. The antibody or antibody fragment thereby recruits Gprl24 via RECK or Reck via Gprl24 to the FZD / LRP complex, thereby assembling the Wnt7-ligand specific RECK / GPR124 / FZD / LRP signalosomes and activating the canonical Wnt7 signaling.

[0151] In another embodiment, the antibody or antibody fragment of the present invention acts by Wnt7-specific RECK / GPR124 / Fzd / LRP-mediated signaling, in which said antibody or antibody fragment binds specifically to RECK and / or Gprl24 in a FZD-dependent and optionally LRP-independent manner, thereby assembling the Wnt7-ligand specific RECK / GPR124 / FZD / LRP signalosomes and activating the canonical Wnt7 signaling.

[0152] Accordingly, in particular embodiments, the antibody or antibody fragment as disclosed herein is able to bind to Fzd, and GPR124 and / or RECK, preferably at a cell membrane.

[0153] The skilled person shall further appreciate that a GPR124 / RECK / FZD / LRP receptor complex, as envisaged herein, may include further component(s), which may or need not functionally modulate the complex. For example, the complex may include Dishevelled (Dvl), forming intracellular scaffolds capable of bridging GPR124 and Fzd.

[0154] In particular embodiments, the antibody or antibody fragment of the invention as disclosed herein induces or has a Wnt signaling activity of at least 35%, preferably at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, preferably 100%, of the GPR124 / RECK / Fzd / LRP-mediated Wnt signaling activity of the full-length wild-type Wnt7 protein. In preferred embodiments, the antibody or antibody fragment as disclosed herein has at least 70% of the GPR124 / RECK / Fzd / LRP-mediated Wnt signaling activity of the full-length wild-type Wnt7 protein. In particular embodiments, the antibody or antibody fragment of the invention as disclosed herein may activate GPR124 / RECK / Fzd / LRP-mediated Wnt signaling in the presence of all four coreceptors more than 30% of the activation by full-length wild-type Wnt7 protein. For example, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 95%, more than 99% and all ranges and subranges therein.

[0155] In one embodiment, the antibody or antibody fragment of the invention comprises one or more Fzd binding domains, and further one or more Gprl24 binding domains and / or Reck binding domains.

[0156] In an embodiment, the antibody or antibody fragment of the application comprises a targeting moiety having an antigen recognition domain that recognizes an epitope presenton Fzd, and further an antigen recognition domain that recognizes an epitope present on Reck and / or an antigen recognition domain that recognizes an epitope present on Gprl24. In an embodiment, the antigen-recognition domain recognizes one or more linear epitopes or one or more conformational epitopes present on Fzd.In an embodiment, the antigen-recognition domain recognizes one or more linear epitopes or one or more conformational epitopes present on Reck and / or on Gprl24.

[0157] In an embodiment, the antibody or antibody fragment herein disclosed can bind to RECK proteins involved in Wnt signaling. RECK is composed of five N-terminal cysteine-knot (CK) motifs or regions (i.e. CK1, CK2, CK3, CK4 and CK5), a cysteine-rich domain (CRD) and three Kazal motifs preceding a Glycosylphosphatidylinositol (GPI)-anchor site. The CK motifs, the CRD and the Kazal motifs are located extracellularly. Accordingly, in embodiments, the antibody or antibody fragment herein disclosed capable of binding to the RECK protein may bind the CK1 motif, CK2 motif, CK3 motif, CK4 motif, CK5 motif, CRD, and / or one or more of the Kazal motifs of the RECK protein, preferably said antibody or antibody fragment binds to the CK4 and / or CK5 regions of the RECK protein.

[0158] Accordingly, in particular embodiments, the antibody or antibody fragment as disclosed herein is capable of binding to the CK4 and / or CK5 regions of RECK polypeptide. The CK4 motif spans from amino acid C216 to C263, and the CK5 motif spans from amino acid C292 to C338 of the amino acid sequence of the human RECK protein annotated under NCBI Genbank accession number NP 066934.1 as disclosed elsewhere herein. Accordingly, the CK4 motif of human RECK comprises, consists essentially of or consists of the amino acid sequence CCDRAEDHACQNACKRILMSKKTEMEIVDGLIEGCKTQPLPQDPLWQC (SEQ ID N°: 49) and the CK5 motif of human RECK comprises, consists essentially of or consists of the amino acid sequence CCSKANTSTCRELCTKLYSMSWGNTQSWQEFDRFCEYNPVEVSMLTC (SEQ ID N°: 50). In particular embodiments, the antibody or antibody fragment as disclosed herein is capable of binding to the amino acid sequence of SEQ ID N°: 49 and / or the amino acid sequence of SEQ ID N°: 50.

[0159] In another or further particular embodiment, the one or more Reck binding domains or targeting moieties bind an amino acid sequence having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a fragment of the Reck protein as depicted in SEQ ID N°: 6. In a further particular embodiment, said amino acid sequence has a length of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or 30 amino acids. In another further particular embodiment, said fragment of the Reck protein is the amino acid sequence between position 23 and 789 or between position 37 and 338 or between position 627 and673 or between position 698 and 752 or between position 753 and 789 of SEQ ID N°: 6.

[0160] In an embodiment, said RECK-binding domain comprises a RECK specific antibody or comprises the RECK-binding region, preferably the variable region sequence or CDRs, of one or more RECK specific antibody or antibodies. Preferably, the RECK specific antibody is directed to the extracellular portion of the RECK polypeptide, such as directed to the CK4 and / or CK5 region(s) of the RECK polypeptide. Nonlimiting examples of RECK antibodies include antibodies which are known in the art and commercially available, such as from Abeam (e.g. ab88249 and ab899l5 specific for human full-length RECK), Cell Signaling Technology (e.g. #3433 specific for human, mouse, rat, monkey RECK (D8C7)), Santa Cruz (e.g. sc-373929 specific for C-terminus of human RECK).

[0161] In particular embodiments, the RECK-binding domain comprises at least one, at least two, at least three, at least four, at least five or at least six CDR(s), each independently having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity with the respective CDR(s) of a RECK specific antibody.

[0162] In particular embodiments, the RECK-binding domain is RECK specific scFv, such as scFv comprising one or more (preferably all 6) CDR(s) of a RECK specific antibody.

[0163] In particular embodiments, the RECK-binding domain is RECK specific VHH, such as VHH comprising one or more (preferably all 3) CDR(s) of a RECK specific heavy chain antibody.

[0164] In particular embodiments, the RECK-binding domain of the antibody or antibody fragment as disclosed herein may comprise two or more CDR(s) each independently having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity with the respective CDR(s) of two or more different RECK specific antibodies. In particular embodiments, said RECK-binding domain of the antibody or antibody fragment as disclosed herein comprises, consists essentially of, consists of or is derived from (e.g., is a biologically active fragment and / or variant of) a RECK-binding polypeptide, such as a Wnt ligand or Wnt polypeptide. Preferably, the RECK-binding domain is derived from a Wnt7 polypeptide (e.g., Wnt7a or Wnt7b) as described elsewhere herein, more preferably from a human or murine Wnt7 polypeptide, even more preferably from a humanWnt7 polypeptide (e.g., human Wnt7a or human Wnt7b). For example, in certain embodiments, the RECK-binding domain may comprise, consist essentially of or consist of a RECK-binding fragment of Wnt7 (e.g., Wnt7a or Wnt7b), preferably of human or mouse Wnt7, more preferably of human Wnt7 (e.g., human Wnt7a or human Wnt7b) or variant thereof In a further example, in certain embodiments, the RECK-binding domain may comprise, consist essentially of or consist of an amino acid sequence having at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity to the amino acid sequence VEPVRASRNKRPTFLKIKKPLSYRKPMDT (SEQ ID N°: 46) or VEVVRASRLRQPTFLRLKQLRSYQKPMET (SEQ ID N°: 48).

[0165] In particular embodiments, said RECK-binding domain is derived from a Wnt7 polypeptide. In particular embodiments, said RECK-binding domain comprises, consists essentially of or consists of an amino acid sequence as set forth in HVEPVRASRNKRPTFLKIKKPLSYRKPMDTDLVYIEKSPNYC (SEQ ID N°: 51), VEPVRASRNKRPTFLKIKKPLSYRKPMDT (SEQ ID N°: 46) or YEWRASRLRQPTFLRIKQLRSYQKPMET (SEQ ID N°: 52), or an amino acid sequence having at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence SEQ ID N°: 51, SEQ ID N°: 46; or SEQ ID N°: 52. In particular embodiments, said RECK-binding domain derived from a Wnt7 polypeptide comprises, consists essentially of or consists of the amino acid sequence SEQ ID N°: 51, SEQ ID N°: 46; or SEQ ID N°: 52.

[0166] In particular embodiments, said RECK-binding domain comprises, consists essentially of, or consists of the amino acid sequence XXXVXAXRXXXXXFLXIXXXXXYXKXXXX (SEQ ID N°: 54), VXAXRXXXXXFLXIXXXXXYXK (SEQ ID N°: 55), XXXVXAXRXXXXXFLXXXXXXXXXKXXXX (SEQ ID N°: 56) or VXAXRXXXXXFLXXXXXXXXXK (SEQ ID N°: 57), wherein X is any amino acid, preferably wherein the amino acid sequence shows at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity to any one of SEQ ID N°: 51, 52 or SEQ ID N°: 46.In an embodiment, said Reck binding domain is chosen from the group of the RECK binding domain of the protein ADAMTS10, eg as depicted in SEQ ID N°: 45 (AA 823-1103 of hADAMTSlO) or a part thereof; the Wnt7a linker region as depicted in SEQ ID N°: 46 or a multitude of said linker regions (eg 3 times the Wnt7a linker); or the extracellular domain of Gprl24 (or part thereof). In an embodiment, said RECK binding domain is derived from the N-terminal extracellular domain of Gprl24, having a sequence showing at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity to SEQ ID N°: 58.

[0167] In an embodiment, said Reck binding domain may comprise, consist essentially of or consist of an amino acid sequence having at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity to the amino acid sequence as given in SEQ ID N°: 45 or SEQ ID N°: 58.

[0168] In another or further embodiment, the antibody or antibody fragment described herein comprises one or more (preferably all 6) CDR(s) of a RECK specific antibody. Said CDRs may be CDRs as described in the table below, and although the CDRs shown below are linked to specific VH and VL sequences, they may be combined as desired.

[0169] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRH1 according to SEQ ID N°: 118, 119, 120, or 121, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 118, 119, 120, or 121.

[0170] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRH2 according to SEQ ID N°: 122, 123, 124, or 125, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 122, 123, 124, or 125.

[0171] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRH3 according to SEQ ID N°: 126, 127, 128, or129, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 126, 127, 128, or 129.

[0172] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRL1 according to SEQ ID N°: 146, 147, 148, or 149, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 146, 147, 148, or 149.

[0173] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRL2 according to SEQ ID N°: 150, 151, 152, or 153, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 150, 151, 152, or 153.

[0174] In an embodiment, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRL3 according to SEQ ID N°: 154, 155, 156, or 157, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 154, 155, 156, or 157.

[0175] In preferred embodiments, the antibody or antibody fragment comprises one or more Reck binding domains comprising a CDRH1, CDRH2, CDRH3; and / or a CDRL1, CDRL2, CDRL3 as combined in the table below.

[0176] In another or further particular embodiment, the one or more Gprl24 binding domains bind an amino acid sequence having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a fragment of the Gprl24 protein as depicted in SEQ ID N°: 7. In a further particular embodiment, said amino acid sequence has a length of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25 or 30 amino acids. In another further particular embodiment, said fragment of the Gprl24 protein is the amino acid sequence between position 34 and 344 or between position 82 and 106 or between position 107 and 130 or between position 131 and 154 or between position 156 and 178 or between position 190 and 240 or between position 247 and 344 of SEQ ID N°: 7.

[0177] In another or further embodiment, the antibody or antibody fragment of the application comprises a full-length multimeric protein that includes two heavy chains and two light chains. Each heavy chain includes one variable region (e.g. VH) and at least three constant regions (e.g. CHI, CH2 and CH3), and each light chain includes one variable region (VL) and one constant region (CL). As described above in thedefinitions section, the variable regions determine the specificity of the antibody and comprise three hypervariable regions also known as complementarity-determining regions (CDRs) that contribute to the antibody binding specificity.

[0178] In an embodiment, said Gprl24 binding domain is derived from RECK, preferably the human RECK sequence. The N-terminal part of RECK was reported to bind to the leucine-rich repeat (LRR) and immunoglobulin (Ig) domains of Gprl24. In an embodiment, said Gprl24 binding domain comprises one or more Cysteine-Knot (CK) domains derived from RECK, preferably human RECK. In an embodiment, said antibody or antibody fragment comprises CK1 to CK5 of RECK, preferably human RECK (see SEQ ID N°: 47). In an embodiment, said GPR124 binding domain comprises a portion of CK1-5 of RECK, preferably hRECK. In an embodiment, said Gprl24 binding domain may comprise, consist essentially of or consist of an amino acid sequence having at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity to the amino acid sequence as given in SEQ ID N°: 47.

[0179] In particular embodiments, the Gprl24-binding domain is or comprises a Gprl24 specific IgG or scFv, such as an IgG or scFv comprising one or more (preferably all 6) CDR(s) of a Gprl24 specific antibody.

[0180] In particular embodiments, the Gprl24-binding domain is or comprises a Gprl24 specific VHH, such as VHH comprising one or more (preferably all 3) CDR(s) of a Gprl24 specific heavy chain antibody.

[0181] In particular embodiments, the Gprl24-binding domain of the antibody or antibody fragment as disclosed herein may comprise two or more CDR(s) each independently having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, preferably 100%, sequence identity with the respective CDR(s) of two or more different Gprl24 specific antibodies.

[0182] In another or further embodiment, the antibody or antibody fragment described herein comprises one or more (preferably all 6) CDR(s) of a Gprl24 specific antibody. Said CDRs may be CDRs as described in the table below, and although the CDRs shown below are linked to specific VH and VL sequences, they may be combined as desired.In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRH1 according to SEQ ID N°: 89 or 90, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 89 or 90.

[0183] In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRH2 according to SEQ ID N°: 91 or 92, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 91 or 92.

[0184] In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRH3 according to SEQ ID N°: 93 or 94, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 93 or 94.

[0185] In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRL1 according to SEQ ID N°: 103 or 104, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 103 or 104.

[0186] In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRL2 according to SEQ ID N°: 105 or 106, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 105 or 106.

[0187] In an embodiment, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRL3 according to SEQ ID N°: 107, 108, or 109, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 107, 108, or 109.

[0188] In preferred embodiments, the antibody or antibody fragment comprises one or more Gprl24 binding domains comprising a CDRH1, CDRH2, CDRH3; and / or a CDRL1, CDRL2, CDRL3 as combined in the table below.

[0189] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRH1 according to SEQ ID N°: 180 or 181, or havingmaximally 1 amino acid difference to a sequence according to SEQ ID N°: 180 or 181.

[0190] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRH2 according to SEQ ID N°: 182, 183, or 184, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 182, 183, or 184.

[0191] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRH3 according to SEQ ID N°: 185 or 186, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 185 or 186.

[0192] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRL1 according to SEQ ID N°: 187 or 188, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 187 or 188.

[0193] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRL2 according to SEQ ID N°: 189 or 190, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 189 or 190.

[0194] In an embodiment, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRL3 according to SEQ ID N°: 191, 192, or 193, or having maximally 1 amino acid difference to a sequence according to SEQ ID N°: 191, 192, or 193.

[0195] In preferred embodiments, the antibody or antibody fragment comprises one or more Fzd binding domains comprising a CDRH1, CDRH2, CDRH3; and / or a CDRL1, CDRL2, CDRL3 as combined in the table below.

[0196] In particular embodiments, at least one of the targeting moieties of the antibody or antibody fragment of the application is a single-domain antibody, such as a VHH. The VHH may be derived from, for example, an organism that produces VHH antibodies such as a camelid, a shark, or the VHH may be a designed VHH. VHHs are antibody-derived therapeutic proteins that contain the unique structural and functional properties of naturally occurring heavy-chain antibodies (see definitionsection above). In some embodiments, the single domain antibody as described herein is an immunoglobulin single variable domain or ISVD. In most particular embodiments, at least one of the targeting moieties of the antibody or antibody fragment is a VHH. In further most particular embodiments, said VHH comprises a single amino acid chain having four "framework regions" and three "complementary determining regions" or CDRs. As used herein, "framework region" refers to a region in the variable domain which is located between the CDRs. As used herein, "complementary determining region" or "CDR" refers to variable regions in VHHs that contains the amino acid sequences capable of specifically binding to antigenic targets. In various embodiments, the antibody or antibody fragment of the application comprises a VHH or ISVD having a variable domain comprising at least one CDR1, CDR2, and / or CDR3 sequence. The CDR3 sequence represents an essential feature of a family of ISVDs, more particularly VHHs. An ISVD family is defined herein as a group of ISVD amino acid sequences with high similarity, or even identical, in the CDR3 sequence. By default, ISVDs belong to the same family when binding to the same target epitope. Variations in an ISVD family may be interesting if expression / stability / affinity / crystallization of a representative of that family is poor, as small deviations like single amino acid mutations occurring within one family may improve these properties.

[0197] In an embodiment, the antibody or antibody fragment of the application comprises an antibody or any antibody fragment herein described such as an IgG, scFv, ISVD, or a VHH, that binds Fzd, Gprl24 or Reck respectively.

[0198] In an embodiment, said the antibody or antibody fragment of the application comprises an antibody or any antibody fragment herein described such as an DVD-Ig (Dual-variable domain immunoglobulin) that binds both Fzd and Gprl24 and / or Reck.

[0199] In the present context, a "Dual-Variable Domain Immunoglobulin" or "DVD-Ig" is a bispecific antibody format comprising two variable domains arranged in tandem within each Fab arm of an immunoglobulin, wherein a first variable domain (VH1 / VL1) is positioned N-terminal to a second variable domain (VH2 / VL2), enabling binding to two distinct target antigens. The DVD-Ig maintains an Fc region, allowing for immune effector functions and extended half-life, and may be engineered to optimize binding affinity, specificity, or sequential antigen engagement.In another or further embodiment, the antibody or antibody fragment described herein comprises an anti-Gprl24 IgG or scFv, comprising:

[0200] - a VH sequence according to SEQ ID N° 63, 65, or 67, and

[0201] - a VL sequence according to SEQ ID N° 64, 66, or 68,

[0202] - the VH and VL optionally separated by a linker sequence,

[0203] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0204] Preferably, the antibody or antibody fragment described herein comprises an anti-Gprl24 IgG or scFv, comprising:

[0205] - a VH sequence according to SEQ ID N° 63 and a VL sequence according to SEQ ID N° 64,

[0206] - a VH sequence according to SEQ ID N°65 and a VL sequence according to SEQ ID N° 66, or

[0207] - a VH sequence according to SEQ ID N° 67 and a VL sequence according to SEQ ID N° 68,

[0208] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0209] Preferably, the antibody or antibody fragment described herein comprises at least combinations of the CDRH1, 2, and 3, and CDRL1, 2, and 3 as described below.

[0210] CD CD CD FR FR FR FR CD CD CD FRL FRL FRL FRL ID VH RH RH RH VL

[0211] Hl H2 H3 H4 RL1 RL2 RL3 1 2 3 4 1 2 3

[0212] 05A10 GV PD RLT QAVLTQP RFS QV ISV PSASGTP GS QVQLQESG QL DTS GQRVTIS KS PGLVKPSET QE RS CSGSSS QA GT LSLTCTVSG SG QF NIGSNYV VLT SAS GSISGSSYY PG SLK YWYQQL QP WY LAI WSWIRQPP NIY LVK Wl LSS PGTAPKL AA PSA QQ SG GRGLEWIG YS PSE RQ VTA WG LIYRNNQ SG WD SG LP LRS NIYYSGSTY GS LLV TLS PP AD QG RPSGVPD SSS RN DS TP GT ED FG YNPSLKSRL TYY QG LTC GR TAV TM RFSGSKS NIG NQ LS GQ AP EA GG TISVDTSRS GS NP VD TVS GL YY VTV GTSASLAI SN RP GV RVT KLL DY TKL QFSLKLSSV SYY SLK AF GG EWI CA SS SGLRSED YVY S V ISC IY YC TVL TAADTAVYY WS S DI SIS G R (SE EADYYCA (SE (SE (SE (SE (SE (SE (SE CARLLVQG (SE (SE (SE (SE (SE (SE Q AWDDSL Q Q Q Q Q Q Q VDAFDIWG Q Q Q Q Q Q ID SGWFG ID ID ID ID ID ID ID QGTMVTVS ID ID ID ID ID ID N°: GGTKLTV N°: N°: N°: N°: N°: N°: N°: S (SEQ ID N°: N°: N°: N°: N°: N°: 102 L (SEQ ID 103 105 107 110 113 115 117 N°: 63) 89) 91) 93) 95) 97) 99) ) N°: 64) ) ) ) ) ) ) ) 06C03 QVQLQESG GS NIY LLV QV Wl RLT WG SYVLTQP SG RN AA SYV WY GV FG PGLVKPSET SYY YS QG QL RQ ISV QG PSASGTP SSS NQ WD LTQ QQ PD GG LSLTCTVSG WS GS VD QE PP DT TM GQRVTIS NIG RP DS PPS LP RFS TKL GSISGSSYY (SE TYY AF SG GK SR VTV CSGSSS SN S LN AS GT GS TVL

[0213]

[0214] WSWIRQPP Q NP DI PG GL NQ SS NIGSNYV YVY (SE GV GT AP KS (SEGKGLEWIG ID SLK (SE LVK EWI FSL (SE YWYQQL (SE Q V PG KLL GS Q NIYYSGSTY N°: S Q PSE G KLS Q PGTAPKL Q ID (SE QR IY SAS ID YNPSLKSRL 89) (SE ID TLS (SE SVT ID LIYRNNQ ID N°: Q VTI (SE LAI N°: TISVDTSRN Q N°: LTC Q AA N°: RPSGVPD N°: 105 ID SC Q SG 117 QFSLKLSSV ID 93) TVS ID DT 102 RFSGSKS 103 ) N°: (SE ID LQ ) TAADTAVYY N°: GG N°: AVY ) GSSASLA ) 108 Q N°: SE CARLLVQG 91) SIS 98) YC ISGLQSE ) ID 113 DE VDAFDIWG (SE AR DEADYYC N°: ) AD QGTMVTVS Q (SE AAWDDS 111 YY S (SEQ ID ID Q LNGWFG ) C N°: 65) N°: ID GGTKLTV (SE 95) N°: L (SEQ ID Q 100 N°: 66) ID ) N°:

[0215] 116 ) 06A05 GV RVT PD QV MS SYELTQP RFS QL VD PSASGTP GS QVQLQESG QE TSK GQRVTIS KS PGLVKPSET SG NQ CSGSSS SYE GT LSLSCTVSG PG FSL NIGTNYV LTQ SAS DSISSRSHY LVK NL YWYQQL PPS WY LAI WGWIRQP TIY PSE Wl SSV PGMAPKL AT AS QQ SG PGKGLEWI YS TLS RQ TAA WG LIYSDDQ SG WD GT LP LRS GTIYYSGNT GN LR LS pp DT QG RPSGVPD SSS SD DS PG GM ED FG YYNPSLKSR SR TYY GS CT GK AVY TM RFSGSKS NIG DQ LS QR AP EA GG VTMSVDTS SH NP VD VS GL FC VTV GTSASLAI TNY RP GV VTI KLL DY TKL KNQFSLNL YW SLK AF GD EWI Al SS SGLRSED VY S V SC IY YC TVL SSVTAADTA G S DI SIS G (SE (SE EADYYCA (SE (SE (SE (SE (SE (SE (SE VYFCAILRG (SE (SE (SE (SE (SE Q Q TWDDSL Q Q Q Q Q Q Q SVDAFDIW Q Q Q Q Q ID ID SGWFG ID ID ID ID ID ID ID GQGTMVTV ID ID ID ID ID N°: N°: GGTKLTV N°: N°: N°: N°: N°: N°: N°: SS (SEQ ID N°: N°: N°: N°: N°: 101 102 L (SEQ ID 104 106 109 112 114 115 117

[0216]

[0217] N°: 67) 90) 92) 94) 96) 98) ) ) N°: 68) ) ) ) ) ) ) ) In another or further embodiment, the antibody or antibody fragment described herein comprises an anti-Reck IgG or scFv, comprising:

[0218] a VH sequence according to SEQ ID N° 69, 71, 73, or 75, and

[0219] - a VL sequence according to SEQ ID N° 70, 72, 74, or 76,

[0220] - the VH and VL optionally separated by a linker sequence,

[0221] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0222] Preferably, the antibody or antibody fragment described herein comprises an anti-Reck IgG or scFv, comprising:

[0223] - a VH sequence according to SEQ ID N° 69 and a VL sequence according to SEQ ID N° 70,

[0224] - a VH sequence according to SEQ ID N° 71 and a VL sequence according to SEQ ID 72,

[0225] - a VH sequence according to SEQ ID N° 73 and a VL sequence according to SEQ ID N° 74, or- a VH sequence according to SEQ ID N° 75 and a VL sequence according to SEQ ID N° 76,

[0226] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0227] Preferably, the antibody or antibody fragment described herein comprises at least combinations of the CDRH1, 2, and 3, and CDRL1, 2, and 3 as described below.

[0228] CD CD CD FR FR FR FR CD CD CD FRL FRL FRL FRL ID VH RH RH RH VL

[0229] Hl H2 H3 H4 RL1 RL2 RL3 1 2 3 4 1 2 3

[0230] 01A08 RLT GV QL MS PS QL VD RFS QLQLQESG QE TSK DIVMTQS GS PGRVKPSE SG NQ PSSLSAS GS TLSLTCKVS PG FSL VGDRVTI DIV GT GFSISRGYY RV KM TCRASQS MT DFT WAWVRQA KPS WV RS ISRYLNW QS WY LTI PGKGLEWI NIH AFY ETL RQ VTA YQQKPG PSS QQ SSL GNIHHSGS HS GS SLT AP AD WG KAPKLLM RA LSA KP QP TYYSPSLKS GS GN CK GK TAV QG YAASSLQ SQ QQ SV GK ED FG RLTMSVDT RG TYY YYV VS GL YF TTV SGVPSRF SIS AA SYS GD AP FAT GG SKNQFSLK YY SPS TAL GF EWI CA IVS SGSGSGT RYL SSL TPL RVT KLL YY TKV MRSVTAAD WA LKS DV SIS G R S DFTLTISS N QS T ITC MY C EIK TAVYFCARA (SE (SE (SE (SE (SE (SE (SE LQPEDFA (SE (SE (SE (SE (SE (SE (SE FYGSGNYY Q Q Q Q Q Q Q TYYCQQS Q Q Q Q Q Q Q VTALDVWG ID ID ID ID ID ID ID YSTPLTF ID ID ID ID ID ID ID QGTTVIVSS N°: N°: N°: N°: N°: N°: N°: GGGTKVE N°: N°: N°: N°: N°: N°: N°: (SEQ ID N0: 118 122 126 130 134 138 142 IK (SEQ 146 150 154 158 162 165 168 69) ) ) ) ) ) ) ) ID N°: 70) ) ) ) ) ) ) ) 01F11 RFT GV EV ISR PS QL DN RFS EVQLVESG VES SN DIQLTQS GS GALVQPGG GG NR PSSLSAS GS SLRLSCAAS ALV LYL VGDRVTI DIQ GT GFTVSSNY AEL QP QM TCRASQ LTQ DFT MSWVRQA VIY LPF GG WV NS GISSYLA SPS WY LTI PGKGLEWV SG RV SLR RQ LRA WYQQKP SLS QQ SG SVIYSGGST GS GV LS AP EDT WG GKAPKLLI RA AS KP LQ YYAGSVKG TYY VIS CA GK GV QG YAASTLQ SQ QQ VG GK AE FG RFTISRDNS SN AG KYY AS GL YY TTV SGVPSRF GIS AA SYS DR AP DF QG NNRLYLQM YM SVK GL GFT EW CA TVS SGSGSGT SYL STL SPI VTI KLL ATY TRL NSLRAEDT S G DV VS VS R S DFTLTISG A QS T TC IY YC DIK GVYYCARA (SE (SE (SE (SE (SE (SE (SE LQAEDFA (SE (SE (SE (SE (SE (SE (SE ELLPFRVGV Q Q Q Q Q Q Q TYYCQQS Q Q Q Q Q Q Q VISKYYGLD ID ID ID ID ID ID ID YSSPITFG ID ID ID ID ID ID ID VWGQGTTV N°: N°: N°: N°: N°: N°: N°: QGTRLDI N°: N°: N°: N°: N°: N°: N°: TVSS (SEQ 119 123 127 131 135 139 143 K (SEQ ID 147 151 155 159 163 166 169 ID N°: 71) ) ) ) ) ) ) ) N°: 72) ) ) ) ) ) ) ) 03A04 EVQLVESG NY SIIA NS EV WV RFT WG DIQMTQS RA AA HQ DIQ WY GV FG GGLVQPG AM SD GR QL RQ ISR QG PSSLSAS SQ SN TYT MT QQ PS QG GSLRLACTT S VST TIF VES TP DN TLV AGDRVTI SIN LQ THT QS KP RFS TKL SGFTFSNY (SE YYA GV GG GG SN TVS TCRASQS TYL S (SE PSS GK GS EIK AMSWVRQ Q DS VMI GL RLE DTL S INTYLNW N (SE Q LSA AP GS (SE

[0231]

[0232] TPGGRLEW ID VR GE VQ WV YL (SE YQQKPG (SE Q ID SA KLL GT QVSSIIASDV N°: G YG PG S QM Q KAPKLLIY Q ID N°: GD IY DFT ID STYYADSVR 120 (SE FD GS (SE HS ID AASNLQS ID N°: 156 RVT (SE LTI N°: GRFTISRDN ) Q S LRL Q LR N°: GVPSRFS N°: 152 ) ITC Q SSL 170 SNDTLYLQ ID (SE AC ID DE 144 GSGSGT 148 ) (SE ID QP ) MHSLRDED N°: Q TTS N°: DT ) DFTLTISS ) Q N°: ED TAVYFCAK 124 ID GFT 136 AVY LQPEDFA ID 163 FAT NSGRTIFGV ) N°: FS ) FC TYYCHQT N°: ) YY VMIGEYGF 128 (SE AK YTTHTFG 160 C DSWGQGT ) Q (SE QGTKLEI ) (SE LVTVSS ID Q K (SEQ ID Q (SEQ ID N0: N°: ID N°: 74) ID 73) 132 N°: N°:

[0233] ) 140 165 ) ) 07B05 RVT GIP QV ISV QSVLTQP DR QVQLVESG QL DM PSVSAAP FTG PGLVKPSQ VES SK GQHVTIS QS SKS TLSLTCTVS GP NQ CSGSSS VLT GT GGSISSGD GL FSL NVGKNF QP SAT YYWSWIRQ YIY VKP Wl KLS VSWYQQ PSV WY LGI HPGKGLE YS SQ RQ SVT LPGTVPK SG SA QQ TGL WIGYIYYSG GS ERL TLS HP AA WG LLIYDNN SSS GT AP LP QT STYHNPSL SG TYH KG LTC GK DT QG RRPSGIP NV DN WD GQ GT GD FG KSRVTISVD DY NP LD TVS GL AVY TM DRFTGSK GK NR TSL HV VPK EA GG MSKNQFSL YW SLK AF GG EWI YC VTV SGTSATL NF RP SA TIS LLI DY TRL KLSSVTAAD S S DI SIS G AR SS GITGLQT VS S W C Y YC TVL TAVYYCARE (SE (SE (SE (SE (SE (SE (SE GDEADYY (SE (SE (SE (SE (SE (SE (SE RLKGLDAF Q Q Q Q Q Q Q CGTWDT Q Q Q Q Q Q Q DIWGQGT ID ID ID ID ID ID ID SLSAWF ID ID ID ID ID ID ID MVTVSS N°: N°: N°: N°: N°: N°: N°: GGGTRLT N°: N°: N°: N°: N°: N°: N°: (SEQ ID N0: 121 125 129 133 137 141 145 VL (SEQ 149 153 157 161 164 167 171

[0234]

[0235] 75) ) ) ) ) ) ) ) ID N°: 76) ) ) ) ) ) ) ) In a preferred embodiment, the antibody or antibody fragment described herein comprises an anti-Gprl24 IgG or scFv, comprising:

[0236] - a VH sequence according to SEQ ID N° 63, 65, or 67, and

[0237] - a VL sequence according to SEQ ID N° 64, 66, or 68,

[0238] - the VH and VL optionally separated by a linker sequence,

[0239] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences,

[0240] and / or an anti-Reck IgG or scFv, comprising:

[0241] - a VH sequence according to SEQ ID N° 69, 71, 73, or 75, and

[0242] a VL sequence according to SEQ ID N° 70, 72, 74, or 76,

[0243] - the VH and VL optionally separated by a linker sequence,

[0244] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0245] In another or further embodiment, the antibody or antibody fragment described herein comprises an anti-Fzd IgG or scFv, comprising:

[0246] a VH sequence according to SEQ ID N° 174, 175, or 176, anda VL sequence according to SEQ ID N° 177, 178, or 179, - the VH and VL optionally separated by a linker sequence,

[0247] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0248] Preferably, the antibody or antibody fragment described herein comprises an anti-Fzd IgG or scFv, comprising:

[0249] - a VH sequence according to SEQ ID N° 174 and a VL sequence according to SEQ ID N° 177,

[0250] - a VH sequence according to SEQ ID N° 175 and a VL sequence according to SEQ ID 178, or

[0251] - a VH sequence according to SEQ ID N° 176 and a VL sequence according to SEQ ID N° 179,

[0252] optionally having 1, 2, 3, 4, or 5 amino acid difference to any one of the FRs in said VH and / or VL sequences.

[0253] Preferably, the antibody or antibody fragment described herein comprises at least combinations of the CDRH1, 2, and 3, and CDRL1, 2, and 3 as described below.

[0254] Anti-Fzd sequences

[0255] ID VH CDR1 CDR CDR3 VL CDR1 CDR CDR3

[0256] 2 2

[0257] 1 EVQLVESGGGLVKPGG FTFTN SAIS CARAT DIQMTQSPSSLSASVGDRVTI RASQ AAS CQQSY SLRLSCAASGFTFTNYA YAMS GSG GFGTV TCRASQSISSYLNWYQQKPG SISSY SLQ STPLTF MSWVRQAPGKGLEWV (SEQ GST VFDY KAPKLLIYAASSLQSGVPSRFS LN S (SEQ SAISGSGGSTYYADSVK ID N°: YYA W GSGSGTDFTLTISSLQPEDFAT (SEQ (SE ID N°: GRFTISRDDSKNTLYLQ 180) (SE (SEQ YYCQQSYSTPLTFGGGTKVEI ID N°: Q ID 191) MNSLKTEDTAVYYCAR Q ID ID N°: KR (SEQ ID N0: 177) 187) N°:

[0258] ATGFGTWFDYWGQG N°: 185) 189) TLVTVSS (SEQ ID N0: 182)

[0259] 174)

[0260] 2 EVQLVESGGGLVQPG NSSFY TVY AFPGS DIQMTQSPSSLSASVGDRVTI SVSSA SAS SSYSLI GSLRLSCAASGFNSSF FM PYL YHPM TCRASQSVSSAVAWYQQKPG (SEQ SLY (SEQ YFMHWVRQAPGKGLE (SEQ DYT (SEQ KAPKLLIYSASSLYSGVPSRFS ID N°: S ID N°: WVATVYPYLDYTYYAD ID N°: Y ID N°: GSRSGTDFTLTISSLQPEDFAT 188) (SE 192) SVKGRFTISADTSKNTA 181) (SE 186) YYCQQSSYSLITFGQGTKVEIK Q ID YLQMNSLRAEDTAVYY Q ID LEDKTHTKV (SEQ ID N0: 178) N°:

[0261] CARAFPGSYHPMDYW N°: 190) GQGTLVTVSS (SEQ ID 183)

[0262] N°: 175)

[0263] 3 EVQLVESGGGLVQPG NSSFY ATV AFPGS DIQMTQSPSSLSASVGDRVTI SVSSA SAS VSSA GSLRLSCAASGFNSSF FM YPY YHPM TCRASQSVSSAVAWYQQKPG (SEQ SLY (SEQ YFMHWVRQAPGKGLE (SEQ LDY (SEQ KAPKLLIYSASSLYSGVPSRFS ID N°: S ID N°: WVATVYPYLDYTYYAD ID N°: TY ID N°: GSRSGTDFTLTISSLQPEDFAT 188) (SE 193) SVKGRFTISADTSKNTA 181) (SE 186) YYCQQVSSATFGQGTKV Q ID YLQMNSLRAEDTAVYY Q ID (SEQ ID N°: 179) N°:

[0264] CARAFPGSYHPMDYW N°: 190) GQGTLVTVSS (SEQ ID 184)

[0265]

[0266] N°: 176)Based on the above, any combination of IgGs, scFvs, antibody formats listed above is deemed disclosed herein. In addition, an although some specific combinations are shown, any combinations of CDRs listed above is deemed disclosed herein.

[0267] Humanization

[0268] In one embodiment, the antibody or antibody fragment of the current application comprises one or more immunoglobulin single variable domains, VHHs, scFvs, or a DVD-Ig, for example targeting Fzd, Gprl24 and / or Reck, that has been "humanized," i.e., one or more amino acid residues in the amino acid sequence of the non-human VHH, scFv, VH, and / or VL domain obtained by immunization is replaced by one or more of the amino acid residues that occur at the corresponding position(s) in a VH and / or VL domain from a conventional 4-chain antibody from a human being to increase the degree of sequence identity with the closest human germline sequence. Potentially useful humanizing substitutions can be ascertained by comparing the sequence of the framework regions of a naturally occurring VHH, VH, or VL domain (e.g., in a scFv or DVD-Ig) with the corresponding framework sequence of one or more closely related human VH and / or VL sequence(s), after which one or more of the potentially useful humanizing substitutions (or combinations thereof) thus determined can be introduced into said sequence (in any manner known per se, as further described herein), and the resulting humanized antibody or fragment can be tested for affinity for the target, for stability, for ease and level of expression, and / or for other desired properties. In this way, by means of a limited degree of trial and error, other suitable humanizing substitutions (or suitable combinations thereof) can be determined by the skilled person. Also, based on what is described before, (the framework regions of) an immunoglobulin single variable domain, such as a VHH domain, or other antibody fragment such as an scFv or a DVD-Ig, may be partially humanized or fully humanized.

[0269] Therefore, in various embodiments, the antibody or antibody fragment of the application comprise a targeting moiety comprising an amino acid sequence having one or more amino acid mutations with respect to the above mentioned sequences. In various embodiments, the antibody or antibody fragment comprises a targeting moiety comprising an amino acid sequence having one, or two, or three, or four, or five, or six, or seven, or eight, or nine, or ten, or fifteen, or twenty amino acid mutations with respect to the above mentioned sequences. In some embodiments, the one or more amino acid mutations may be independently selected from substitutions, insertions, deletions, and truncations. In some embodiments, theamino acid mutations are amino acid substitutions, and may include conservative and / or non-conservative substitutions. In particular embodiments, the one or more amino acid mutations may be in the CDRs of the targeting moiety (e.g., the CDR1, CDR2 or CDR3 regions). In other particular embodiments, the one or more amino acid mutations may be in the framework regions of the targeting moiety (e.g., the FR1, FR2, FR3, or FR4 regions). In most particular embodiments, said one or more amino acid mutations are only present in the framework regions of said FRo binding agents. "Conservative substitutions" may be made, for instance, on the basis of similarity in polarity, charge, size, solubility, hydrophobicity, hydrophilicity, and / or the amphipathic nature of the amino acid residues involved. The 20 naturally occurring amino acids can be grouped into the following six standard amino acid groups: (1) hydrophobic: Met, Ala, Vai, Leu, He; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. As used herein, "conservative substitutions" are defined as exchanges of an amino acid by another amino acid listed within the same group of the six standard amino acid groups shown above. For example, the exchange of Asp by Glu retains one negative charge in the so modified polypeptide. In addition, glycine and proline may be substituted for one another based on their ability to disrupt o-helices. As used herein, "non-conservative substitutions" are defined as exchanges of an amino acid by another amino acid listed in a different group of the six standard amino acid groups (1) to (6) shown above. In various embodiments, the substitutions may also include non-classical amino acids (e.g. selenocysteine, pyrrolysine, N-formylmethionine 0-alanine, GABA and 6-Aminolevulinic acid, 4-aminobenzoic acid (PABA), D-isomers of the common amino acids, 2,4-diaminobutyric acid, o-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, y-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosme, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, 0-alanine, fluoro-amino acids, designer amino acids such as [3 methyl amino acids, C o-methyl amino acids, N o-methyl amino acids, and amino acid analogs in general).

[0270] Humanization can be performed using humanization techniques known in the art. In some embodiments, possible humanizing substitutions or combinations of humanizing substitutions may be determined by methods known in the art, for example without the purpose of being limiting, by a comparison between the sequence of a antibody fragment and the sequence of a naturally occurring human sequence. In some embodiments, the humanizing substitutions are chosen such thatthe resulting humanized antibody fragment still retain advantageous functional properties. Generally, as a result of humanization, the antibody fragment of the application may become more "human-like", while still retaining favourable properties such as a reduced immunogenicity, compared to the corresponding naturally occurring antibody fragment domains. In various embodiments, the humanized antibody fragments of the application can be obtained in any suitable manner known in the art and thus are not strictly limited to polypeptides that have been obtained using a polypeptide that comprises a naturally occurring antibody fragment domain as a starting material. Indeed, modification of the amino acid sequences may be achieved using any known technique in the art e.g., site-directed mutagenesis or PCR based mutagenesis. Such techniques are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Plainview, N.Y., 1989 and Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, New York, N.Y., 1989.

[0271] In various embodiments, the mutations that were introduced for example to humanize antibody or antibody fragment do not substantially reduce the present antibody or antibody fragments capability to specifically bind to the human Fzd, Gprl24 and / or Reck and to agonize the Gprl24 / Reck / Fzd / LRP5 or LRP6-mediated Wnt signaling.

[0272] Methods of production

[0273] The antibody or antibody fragment of the application are not limited to a specific biological source or to a specific method of preparation. Methods for producing the antibody or antibody fragment of the application are described herein. For example, DNA sequences encoding the antibody or antibody fragment of the application can be easily prepared by the art-known techniques such as cloning, hybridization screening and Polymerase Chain Reaction (PCR). Standard techniques for cloning, DNA isolation, amplification and purification, for enzymatic reactions involving DNA ligase, DNA polymerase, restriction endonucleases and the like, and various separation techniques are those known and commonly employed by those skilled in the art. A number of standard techniques are described in Sambrook et al. (1989), Maniatis et al. (1982), Wu (ed.) (1993) and Ausubel et al. (1992). Alternatively, DNA sequences encoding the antibody or antibody fragment of the application can be chemically synthesized using methods known in the art. Synthetic DNA sequences can be ligated to other appropriate nucleotide sequences, including for example expression control sequences, to produce gene expression constructs encoding the desired FRo antibody or antibody fragment.Accordingly, in various embodiments, the present application provides for isolated nucleic acids comprising a nucleotide sequence encoding the antibody or antibody fragment described in current application for example, encoding the proteins disclosed herein and formulated for vectorized gene therapy delivery for in vivo or ex vivo expression, and, where the coding sequence exceeds a single-vector packaging capacity (e.g., AAV), employing dual-vector strategies such as dual AAV delivery.

[0274] Dual AAV," as used herein, refers to a two-vector adeno-associated virus delivery strategy in which complementary portions of a coding sequence and / or expression cassette are packaged separately into two AAV genomes and co-administered such that, following infection of the same target cell, the full-length expression unit is reconstituted (e.g., by homologous overlap / concatemerization, 5' / 3' trans-splicing, and / or split-intein-mediated protein reconstitution), thereby enabling expression of transgenes exceeding the packaging capacity of a single AAV vector. One embodiment further discloses an expression cassette comprising said nucleic acid molecule. More specific embodiments disclose the expression cassette wherein elements for cell- or tissue-specific expression are present e.g., promoters and / or enhancers conferring brain or blood-brain barrier endothelial cell specificity. Further embodiments relate to a vector comprising said expression cassette or said nucleic acid molecule. More particular, said vector may be a viral vector or lentiviral vector, preferably a viral vector specifically directed towards the central and / or peripheral nervous system (e.g., a brain-specific viral vector). In further particular embodiments, the viral vector is a central nervous system (CNS) neuron-specific adeno-associated virus serotype 9 (AAV9) mutant. In preferred embodiments, the viral vector is a blood brain barrier endothelial cell-specific viral vector. In further preferred embodiments, the viral vector is a blood brain barrier endothelial cellspecific capsid adeno-associated virus serotype 2 (AAV2) mutant. This may be configured as a single- or dual-AAV system for delivery of larger transgenes; in certain embodiments, reconstitution of the full-length coding sequence in target cells is achieved using overlapping genomes, 5' / 3' trans-splicing, and / or split-intein-mediated protein reconstitution. In preferred embodiments, the viral vector is a blood retina barrier endothelial cell-specific viral vector.

[0275] Factors of importance in selecting a particular vector include inter alia: choice of recipient cell, ease with which recipient cells that contain the vector may be recognized and selected from those recipient cells which do not contain the vector; the number of copies of the vector which are desired in particular recipient cells; whether it is desired for the vector to integrate into the chromosome or to remainextra-chromosomal in the recipient cells; and whether it is desirable to be able to "shuttle" the vector between recipient cells of different species, as well as the packaging capacity of the vector and whether single- or multi-vector (e.g., dual AAV) delivery is indicated by transgene size.

[0276] Expression vectors can be autonomous or integrative. A nucleic acid can be introduced into a cell in the form of an expression vector such as a plasmid, phage, transposon, cosmid or virus particle. The recombinant nucleic acid can be maintained extra-chromosomally or it can be integrated into the cell chromosomal DNA. Expression vectors can contain selection marker genes encoding proteins required for cell viability under selected conditions (e.g., URA3, which encodes an enzyme necessary for uracil biosynthesis, or LEU2, which encodes an enzyme required for leucine biosynthesis, or TRP1, which encodes an enzyme required for tryptophan biosynthesis) to permit detection and / or selection of those cells transformed with the desired nucleic acids. Expression vectors can also include an autonomous replication sequence (ARS). The ARS may comprise a centromere (CEN) and an origin of replication (ORI). For example, the ARS may be ARS18 or ARS68.

[0277] Integrative vectors generally include a serially arranged sequence of at least a first insertable DNA fragment, a selectable marker gene, and a second insertable DNA fragment. The first and second insertable DNA fragments are each about 200 (e.g., about 250, about 300, about 350, about 400, about 450, about 500, or about 1000 or more) nucleotides in length and have nucleotide sequences which are homologous to portions of the genomic DNA of the cell species to be transformed. A nucleotide sequence containing a nucleic acid of interest for expression is inserted in this vector between the first and second insertable DNA fragments, whether before or after the marker gene. Integrative vectors can be linearized prior to transformation to facilitate the integration of the nucleotide sequence of interest into the cell genome. Prior to introducing the vectors into a cell of interest, the vectors can be grown (e.g., amplified) in bacterial cells such as Escherichia coli (E. coli). The vector DNA can be isolated from bacterial cells by any of the methods known in the art, which result in the purification of vector DNA from the bacterial milieu. The purified vector DNA can be extracted extensively with phenol, chloroform, and ether, to ensure that no E. coli proteins are present in the plasmid DNA preparation, since these proteins can be toxic to mammalian cells, and, when used for vectorized gene therapy delivery, the resulting vector preparations can be formulated for single- or dual-AAV administration as described herein.In order to produce the antibodies or antibody fragments of current application, expression vectors comprising a nucleic acid sequence encoding said antibodies or antibody fragments can then be introduced into host cells through transfection, transformation, or transduction techniques. Hence, in various embodiments, the present application provides for a host cell comprising a nucleic acid encoding one of the antibodies or antibody fragments of the present application. For example, nucleic acids encoding the antibodies or antibody fragments of the application can be introduced into host cells by retroviral transduction. Illustrative host cells are E.coli cells, Chinese hamster ovary (CHO) cells, yeast cells such as Pichia spp., human embryonic kidney 293 (HEK 293) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and myeloma cells. Transformed host cells can be grown under conditions that permit the host cells to express the genes that encode the antibodies or antibody fragments of the application.

[0278] Following expression, the antibodies or antibody fragments can be harvested and purified using techniques well known in the art, e.g. affinity tags such as glutathione-S-transferase (GST) and histidine (His) tags or by chromatography. Specific expression and purification conditions will vary depending upon the expression system employed. For example, if a gene is to be expressed in E. coli, it is first cloned into an expression vector by positioning the engineered gene downstream from a suitable bacterial promoter, e.g. Trp or Tac, and a prokaryotic signal sequence. In another example, if the engineered gene is to be expressed in eukaryotic host cells, e.g. CHO cells, it is first inserted into an expression vector containing for example, a suitable eukaryotic promoter, a secretion signal, enhancers, and various introns. In an embodiment, the antibody or antibody fragment of the application comprises a His tag, a FLAG-tag and / or a Myc tag. In an embodiment, the antibodies or antibody fragments of the application comprises a His tag and a proteolytic site to allow cleavage of the His tag.

[0279] Current application thus also provides a host cell comprising one of the antibodies or antibody fragments described herein. Host cells comprising one of the nucleic acid molecules or the expression cassettes or the vectors of the application are provided herein as well. Host cells can be either prokaryotic or eukaryotic. Representative host cells that may be used with the invention include, but are not limited to, bacterial cells, yeast cells, plant cells and animal cells. Bacterial host cells suitable for use with the invention include Escherichia spp. cells, Bacillus spp. cells, Streptomyces spp. cells, Erwinia spp. cells, Klebsiella spp. cells, Serratia spp. cells, Pseudomonas spp. cells, and Salmonella spp. cells. Yeast host cells suitable for usewith the invention include species within Saccharomyces, Schizosaccharomyces, Kluyveromyces, Pichia (e.g. Pichia pastoris), Hansenula (e.g. Hansenula polymorpha), Yarowia, Schwaniomyces, Schizosaccharomyces, Zygosaccharomyces and the like. Saccharomyces cerevisiae, S. carlsbergensis and K. lactis are the most commonly used yeast hosts and are convenient fungal hosts. Animal host cells suitable for use with the invention include insect cells and mammalian cells (most particularly derived from Chinese hamster (e.g. CHO), and human cell lines, such as HeLa). Exemplary insect cell lines include, but are not limited to, Sf9 cells, baculovirus-insect cell systems (e.g. review Jarvis 2003 Virology 310:1-7). Nonlimiting examples of plant cells include tobacco cells, Arabidopsis cells, tomato cells, maize cells, algae cells, among others. The host cells may be provided in suspension or flask cultures, tissue cultures, organ cultures and the like. Alternatively, the host cells may also be transgenic animals.

[0280] Animal or mammalian host cells suitable for harboring, expressing, and producing one of the FRo binding agents of the application include Chinese hamster ovary cells (CHO), such as CHO-K1 (ATCC CCL-61), DG44 (Chasin et al 1986 Som Cell Mol Genet 12:555-556; Kolkekar et al 1997 Biochemistry 36:10901-10909), CHO-K1 Tet-On cell line (Clontech), CHO designated ECACC 85050302 (CAMR, Salisbury, Wiltshire, UK), CHO clone 13 (GEIMG, Genova, IT), CHO clone B (GEIMG, Genova, IT), CHO-Kl / SF designated ECACC 93061607 (CAMR, Salisbury, Wiltshire, UK), RR-CHOK1 designated ECACC 92052129 (CAMR, Salisbury, Wiltshire, UK), dihydrofolate reductase negative CHO cells (CHO / -DHFR, Urlaub & Chasin 1980 PNAS 77:4216), and dpl2.CHO cells (U.S. Pat. No. 5,721,121); monkey kidney CV1 cells transformed by SV40 (COS cells, COS-7, ATCC CRL-1651); human embryonic kidney cells (e.g., 293 cells, or 293T cells, or 293 cells subcloned for growth in suspension culture, Graham et al 1977 J Gen Virol 36:59, or GnTI KO HEK293S cells, Reeves et al 2002 PNAS 99:13419); baby hamster kidney cells (BHK, ATCC CCL-10); monkey kidney cells (CV1, ATCC CCL-70); African green monkey kidney cells (VERO-76, ATCC CRL-1587; VERO, ATCC CCL-81); mouse sertoli cells (TM4, Mather 1980 Biol Reprod 23:243-251); human cervical carcinoma cells (HELA, ATCC CCL-2); canine kidney cells (MDCK, ATCC CCL-34); human lung cells (W138, ATCC CCL-75); human hepatoma cells (HEP-G2, HB 8065); mouse mammary tumor cells (MMT 060562, ATCC CCL-51); buffalo rat liver cells (BRL 3A, ATCC CRL-1442); TRI cells (Mather, 1982, Annals NYAcad. Sci., 383:44-68); MCR 5 cells; FS4 cells. According to a particular embodiment, the cells are mammalian cells selected from Hek293 cells or COS cells.The host cells described above can be transiently or stably transfected. Such transfection of DNA, such as nucleic acid molecules, expression cassettes or expression vectors into prokaryotic and eukaryotic cells can be accomplished via any technique known in the art, including but not limited to standard bacterial transformations, calcium phosphate co-precipitation, electroporation, or liposome mediated-, DEAE dextran mediated-, polycationic mediated-, or viral mediated transfection. For all standard techniques see, for example, Molecular Cloning: A Laboratory Manual (Sambrook, et al., 1989, Cold Spring Harbor Laboratory Press; Culture of Animal Cells: A Manual of Basic Technique, 2* Ed. (R.I. Freshney. 1987. Liss, Inc. New York, N.Y.). The host cell may also be a recombinant host cell, which involves a cell which has been genetically modified to contain an isolated DNA molecule, nucleic acid molecule or expression construct or vector of the invention. The DNA can be introduced by any means known to the art which are appropriate for the particular type of cell, including without limitation, transformation, lipofection, electroporation or viral mediated transduction.

[0281] Furthermore, in alternative embodiments, the use of the herein described nucleic acid molecules, expression cassettes, or vectors encoding the antibodies or antibody fragments of the application, are provided for the production of antibodies or antibody fragments. In a particular embodiment, said use is provided for production of an intrabody. An intracellular antibody or "intrabody" is an antibody or a fragment of an antibody that is heterologously expressed within a designated intracellular compartment, a process which is made possible through the in-frame incorporation of intracellular trafficking signals. An intrabody can be expressed in any shape or form such as an intact IgG molecule or a Fab fragment, more particularly as genetically engineered antibody fragment for example as single domain intrabodies or VHHs. For a review see Zhu, and Marasco, 2008 (Therapeutic Antibodies. Handbook of Experimental Pharmacology 181. Ed. Springer-Verlag Berlin Heidelberg).

[0282] Medical applications

[0283] As described earlier, the atypical Wnt7 signaling mediated by the Gprl24 / Reck / Fzd / LRP5 or LRP6 complex is of crucial importance to initiate bloodbrain-barrier differentiation and maintenance of BBB function, and blood-retina-barrier differentiation and maintenance of BRB function. Moreover, stimulating Wnt7 signaling in cerebral / retinal endothelial cells with impaired BBB / BRB can restore the BBB / BRB and ameliorate associated disease effects.Therefore, in another aspect of the invention, any antibody or antibody fragment herein disclosed, but also any nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector, or vector herein described is provided for use as a medicament. This is equivalent to providing a method of treatment, comprising the step of administering any antibody or antibody fragment herein disclosed, or any nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector, or vector herein described to a subject or patient in need thereof.

[0284] In particular embodiments, the method of treatment is a method of treatment or prevention of a disorder presenting an abnormal Wnt signaling in RECK and / or GPR124 expressing cells, tissues and / or organs.

[0285] In a particular embodiment, said antibody or antibody fragment, nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector or vector is provided for use in gene therapy, more particularly for use to treat blood-brain barrier integrity.

[0286] In another particular embodiment, said antibody or antibody fragment, nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector or vector is provided for use in gene therapy, more particularly for use in blood-brain barrier endothelial cell-directed gene therapy.

[0287] Accordingly, also provided herein is a method for gene therapy, in particular central and / or peripheral nervous system-directed gene therapy, in a subject in need of said gene therapy comprising: introducing in the subject, in particular in the central and / or peripheral nervous system of the subject, a nucleic acid expression cassette or a vector as described herein; and expressing a therapeutically effective amount of the antibody or antibody fragment encoded by the nucleic acid as taught herein in the subject, in particular the central and / or peripheral nervous system of the subject.

[0288] In particular embodiments, the antibody or antibody fragment or the nucleic acids encoding said antibodies as taught herein are used in mRNA therapy, in particular in blood brain barrier endothelial cell-directed mRNA therapy.

[0289] Accordingly, also provided herein is a method for RNA therapy, preferably mRNA therapy, in particular central and / or peripheral nervous system-directed mRNA therapy, in a subject in need of said mRNA therapy comprising: introducing in the subject, in particular in the central and / or peripheral nervous system of the subject,a nucleic acid encoding any of the herein disclosed antibody or antibody fragment; and expressing a therapeutically effective amount of the antibody or antibody fragment encoded by the nucleic acid as taught herein in the subject, in particular the central and / or peripheral nervous system of the subject.

[0290] An advantage of the use of RNAs therapy, it is generally believed that RNAs do not integrate into the genome and therefore do not have the risk of insertional mutagenesis.

[0291] Any other well-known methods of introducing nucleic acids into animal cells may be used herein. At the simplest, the nucleic acid can be directly injected into the target cell I target tissue. Other methods include fusion of the recipient cell with bacterial protoplasts containing the nucleic acid, the use of compositions like calcium chloride, rubidium chloride, lithium chloride, calcium phosphate, DEAE dextran, cationic lipids or liposomes or methods like receptor-mediated endocytosis, biolistic particle bombardment ("gene gun" method), infection with viral vectors (i.e. derived from lentivirus, adeno-associated virus (AAV), adenovirus, retrovirus or antiviruses), electroporation, and the like. Other techniques or methods which are suitable for delivering nucleic acid (NA) molecules to target cells include the continuous delivery of an NA molecule from poly (lactic-Co-Glycolic Acid) polymeric microspheres or the direct injection of protected (stabilized) NA molecule(s) into micropumps delivering the product. Another possibility is the use of implantable drug-releasing biodegradable microspheres. Also envisaged is encapsulation of NA or providing NA in various types of liposomes (immunoliposomes, PEGylated (immuno) liposomes), cationic lipids and polymers, nanoparticles or dendrimers, poly (lactic-Co-Glycolic Acid) polymeric microspheres, implantable drug-releasing biodegradable microspheres, etc.; and co-injection of NA with protective agent like the nuclease inhibitor aurintricarboxylic acid. It shall be clear that also a combination of different above-mentioned delivery modes or methods may be used.

[0292] In particular embodiments, the antibody or antibody fragment or nucleic acid encoding it is provided in carriers, such as liposomes, lipid nanoparticles, nanostructured lipid carriers, nanoemulsions, polymer nanoparticles, polymer micelle or dendrimers. In a preferred embodiment, said carriers are liposomes or lipid nanoparticles (LNPs).

[0293] In an embodiment, said carriers are lipid-based carriers. Said lipid-based carriers comprise one or more lipids. The one or more lipids can be in solid and / or liquidform. Said lipid-based carriers may be LNPs, lipoplexes, liposomes, phospholipid micelles, solid lipid nanoparticles, nanostructured lipid carriers or nano-emulsions. Lipid-based carriers useful according to the invention include, for example, cationic lipids, liposomes, in particular cationic liposomes, and micelles, and nanoparticles. Cationic lipids may form complexes with negatively charged nucleic acids. Any cationic lipid may be used according to the invention. Liposomes are phospholipid and cholesterol self-assembled bilayer membranes that enclose an aqueous core, where hydrophilic molecules can be incorporated. Hydrophobic compounds can also be incorporated in the lipid bilayer. Liposomes can be classified in (i) small unilamellar vesicles (SUVs); (ii) large unilamellar vesicles (LUVs) and (iii) multilamellar vesicles (MLVs), according to their size and lamellarity. Solid lipid nanoparticles (SLNs) have a spherical shape with an average diameter of 10-1000 nm. They are used as a colloidal NP drug delivery system in which lipid drug carrier solidifies at room temperature as well as at body temperature. Different solid lipids can be exploited to produce SLNs, such as, tripalmitin, cetyl alcohol, cetyl palmitate, glyceryl monostearate, trimyristin, tristearin, stearic acid, etc. SLNs comprise of solid lipid, such as triglycerides, fatty acids, waxes, partial glycerides, and polyethylene glycosylated lipid; emulsifiers, such as polysorbates, poloxamer and lecithin; and water. Nanostructured lipid carriers (NLC), comprise a blend of solid and liquid lipids which results in a partially crystallized lipid system and many have advantages such as enhanced drug loading capacity, drug release modulation flexibility and improved stability.

[0294] In an embodiment, said lipid-based carrier is a lipid nanoparticle. Solid lipid nanoparticles (SLNs, sLNPs), or lipid nanoparticles (LNPs), are nanoparticles composed of lipids that are suited to be used as a drug delivery vehicle for drug compounds, especially polynucleotides such as RNA or DNA.

[0295] In another embodiment, any antibody or antibody fragment herein disclosed, but also any nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector, or vector herein described is provided for use in the treatment of a neurological disorder, and / or in the treatment of an ophthalmological disorder.

[0296] Also provided is a method of treating blood-brain-barrier dysfunction or treating a neurological disorder, or treating blood-retina-barrier dysfunction or treating an ophthalmological disorder such as a retinal vascular disorder or retinal degenerative disorder comprising the step of administering any antibody or antibody fragment herein disclosed, or any nucleic acid, nucleic acid sequence, nucleic acid molecule,expression vector, or vector herein described to a subject or patient in need thereof. In a particular embodiment, said administration is performed by gene therapy, more particularly by blood-brain barrier endothelial cell-directed gene therapy or bloodretina barrier endothelial cell-directed gene therapy.

[0297] Also provided is a method of stimulating, activating or agonizing the Gprl24 / Reck / Fzd / LRP5 or LRP6 mediated Wnt7 signaling in a cell, comprising the step of administering a therapeutically effective amount of any antibody or antibody fragment, nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector, or vector herein described to said cell, wherein said cell expresses at least Gprl24, Reck, Fzd, more particular Fzdl and a LRP protein selected from LRP5 and LRP6. In a further embodiment, said antibody or antibody fragment, nucleic acid, nucleic acid sequence, nucleic acid molecule, expression vector, or vector cannot activate the canonical Wnt signaling or Wnt7 signaling in the absence of RECK and / or GPR124.

[0298] In a particular embodiment, a neurological disorder as used herein is a disorder selected from the list consisting of ischemic stroke, hemorrhagic stroke, ischemia / reperfusion injury, brain aneurysms, arteriovenous malformations (AVMs), cavernous malformations, vasculitis, cerebral hemorrhage, subarachnoid hemorrhage, spinal vascular malformations, carotid artery stenosis, Moyamoya disease intracranial atherosclerosis, and combinations thereof, or multiple sclerosis, brain cancer, glioblastoma, human monogenic neurological disorders, neurodegenerative disorders, dementia, vascular dementia, HIV-l-associated dementia, Alzheimer's disease, Parkinson's disease, Huntington disease, amyotrophic lateral sclerosis, Charcot-Marie Tooth disease, dystonia, infectious brain diseases, traumatic brain or retinal injuries, migraine, neuroinflammation, COVID-19, and chronic traumatic encephalopathy and combinations thereof.

[0299] "Neuroinflammation" can refer to a neurovascular inflammation. Further, neuroinflammation can be triggered by different factors. For example, in non-limiting embodiments neuroinflammation is caused by an injury such as traumatic brain injury and / or spinal cord injury, an infection such as viral or bacterial or fungal infection, exposure to a toxin or due to toxic metabolites, neurodegenerative disease, autoimmune disease, smoking or passive smoking, aging or any combination thereof.Neuroinflammation can be caused by an infectious disease. Said infection may be of bacterial, fungal, parasitic or viral origin. In some instances, the infectious disease is a viral infection. In certain instances, the viral infection comprises infection by one or more of Herpesviridae (Herpes Zoster Virus (VZV) infection, Herpes Simplex Virus (HSV-1 or HSV-2), cytomegalovirus (CMV)), Paramoxyviridae, coronaviruses (SARS-CoV-1 or MERS-CoV or SARS-CoV-2, HCoVNL63, HCoV-229E, HCoV-OC43, or HKU1 or a variant thereof), influenza viruses (influenza virus A, group 1 (H1N1), group 2 (H2N2), group 3 (H3N2), group 5 (H5N1, H5N2, H5N8) or group 7 (H7N7, H7N9), influenza B, influenza C), Zika virus, Japanese encephalitis virus, Epstein-Barr virus, Hepatitis B virus, Hepatitis C virus, Human immunodeficiency virus type 1 (HIV-1), Human papilloma virus (HPV), Human T-cell lymphotropic virus type I (HTLV-1) and Kaposi sarcoma herpesvirus (KSHV).

[0300] Neuroinflammation can be associated with lung condition such as, acute lung injury (ALI) and / or acute respiratory distress syndrome (ARDS), possibly associated with or arising from ventilator use, viral infection, sepsis, or systemic bacterial infections in a subject in need thereof have been established. Said lung condition can be caused by an infectious disease caused by bacterial, fungal, parasitic or viral infection. Preferably said infection is an infection causing respiratory problems or is a respiratory tract infection. Preferably said infection is a viral infection.

[0301] Neuroinflammation can be associated with a bacterial infection. Such inflammation is mostly caused by blood-borne bacteria that cross the blood-brain barrier (BBB) and finally invade the brain parenchyma. Pathogens such as Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae are the main etiological causes of bacterial meningitis.

[0302] In the present context, prevention and / or treatment of neuroinflammation may include encephalitis. In certain instances, the encephalitis is viral encephalitis, such as COVID-19 associated encephalitis. Neuroinflammation may also be related to inflammation of the optic nerve. Neuroinflammation may also be related to acute disseminated encephalomyelitis, such as COVID-19 associated acute disseminated encephalomyelitis. Neuroinflammation may also be related to inflammation of the vitreous retinal ganglion cells.

[0303] In another or further embodiment, the ophthalmological disease or disorder is a retinal vascular disorder or retinal degenerative disorder, preferably said ophthalmological disease or disorder is selected from the group of: retinopathy,retinal vascular disorders such as Norrie disease, diabetic retinopathy, macular degeneration such as wet age-related macular degeneration and diabetic macular edema, familial exudative vitreoretinopathy, osteoporosis-pseudoglioma syndrome, retinal vein occlusion (RVO) including Central Retinal Vein Occlusion (CRVO) and Branch Retinal Vein Occlusion (BRVO), impaired vision due to myopic choroidal neovascularisation, retinopathy of prematurity (ROP), Proliferative Diabetic Retinopathy (PDR), Macular Edema secondary to uveitis, Radiation Retinopathy, Choroidal Neovascularization (CNV) from other causes (including angioid streaks, inflammatory causes such as uveitis and multifocal choroiditis) and traumatic optic neuropathy. The diseases or disorders may for instance also include degenerative ophthalmic diseases, including, without limitation, dry age-related macular degeneration (dry AMD; including geographic atrophy), glaucoma (e.g., primary open-angle glaucoma), retinitis pigmentosa and other inherited retinal dystrophies (e.g., Stargardt disease, Leber congenital amaurosis, cone-rod dystrophy, choroideremia, Best disease), and optic atrophy.

[0304] In embodiments, the antibodies or antibody fragments here described and compositions comprising them are particularly suitable to be administered to a patient prone to develop or suffering from such disorder. In particular embodiments, the antibody or antibody fragment or composition is administered orally or parenterally, e.g., intravenously, intraperitoneally, subcutaneously, or intraocularly.

[0305] In particular embodiments, the pharmaceutical composition as taught herein (see below) is administered to the subject by the injection (e.g., intravenously or intraocularly) or transplantation of allogeneic cells transformed with the vector comprising the nucleic acid or the nucleic acid expression cassette as taught herein. When administered, the injected or transplanted allogenic cells will transcribe and translate the nucleic acid encoding the antibody or antibody fragment as taught herein in vivo.

[0306] The dosage or amount of the antibody or antibody fragment as taught herein, optionally in combination with one or more other active compounds to be administered, depends on the individual case and is, as is customary, to be adapted to the individual circumstances to achieve an optimum effect. Thus, the unit dose and regimen depend on the nature and the severity of the disorder to be treated, and also on factors such as the species of the subject, the sex, age, body weight, general health, diet, mode and time of administration, immune status, and individual responsiveness of the human or animal to be treated, efficacy, metabolic stabilityand duration of action of the compounds used, on whether the therapy is acute or chronic or prophylactic, or on whether other active compounds are administered in addition to the antibody or antibody fragment described in any of the embodiments above. In order to optimize therapeutic efficacy, the antibody or antibody fragment as taught herein can be first administered at different dosing regimens. Typically, levels of the antibody or antibody fragment in a tissue can be monitored using appropriate screening assays as part of a clinical testing procedure, e.g., to determine the efficacy of a given treatment regimen. The frequency of dosing is within the skills and clinical judgement of medical practitioners (e.g., doctors, veterinarians or nurses). Typically, the administration regime is established by clinical trials which may establish optimal administration parameters. However, the practitioner may vary such administration regimes according to the one or more of the aforementioned factors, e.g., subject's age, health, weight, sex and medical status. The frequency of dosing can be varied depending on whether the treatment is prophylactic or therapeutic.

[0307] Toxicity and therapeutic efficacy of the antibody or antibody fragment as described herein or pharmaceutical composition of the invention as described in any of the embodiments, comprising the same can be determined by known pharmaceutical procedures in, for example, cell cultures or experimental animals. These procedures can be used, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50 / ED50. Pharmaceutical compositions that exhibit high therapeutic indices are preferred. While pharmaceutical compositions that exhibit toxic side effects can be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to normal cells (e.g., non-target cells) and, thereby, reduce side effects.

[0308] The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosages for use in appropriate subjects. The dosage of such pharmaceutical compositions lies generally within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For a pharmaceutical composition used as described herein, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasmaconcentration range that includes the IC50 (i.e., the concentration of the pharmaceutical composition which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography.

[0309] In particular embodiments, the antibody or antibody fragment as taught herein is administered using a sustained delivery system, such as a (partly) implanted sustained delivery system. Skilled person will understand that such a sustained delivery system may comprise a reservoir for holding the antibody or antibody fragment as taught herein, a pump and infusion means (e.g., a tubing system). For example, the sustained delivery system may be a mini-osmotic pump system implanted in the brain.

[0310] In particular embodiment, the antibody or antibody fragment as disclosed herein is the main or only active ingredient of the pharmaceutical composition.

[0311] In another embodiment, the pharmaceutical composition as described in any of the embodiments is combined with a second therapy, preferably chosen from surgery, chemotherapy, radiotherapy or immunotherapy.

[0312] Pharmaceutical Compositions and Formulations

[0313] Also disclosed herein are pharmaceutical compositions comprising any of antibody or antibody fragment, nucleic acid, nucleic acid expression cassette, or vector as described herein in any of the embodiments, with an optional pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0314] The term "pharmaceutically acceptable" as used herein is consistent with the art and means compatible with the other ingredients of a pharmaceutical composition and not deleterious to the recipient thereof.

[0315] As used herein, "carrier" or "excipient" includes any and all solvents, diluents, buffers (such as, e.g., neutral buffered saline or phosphate buffered saline), solubilisers, colloids, dispersion media, vehicles, fillers, chelating agents (such as, e.g., EDTA or glutathione), amino acids (such as, e.g., glycine), proteins, disintegrants, binders, lubricants, wetting agents, emulsifiers, sweeteners, colorants, flavourings, aromatisers, thickeners, agents for achieving a depot effect, coatings, antifungal agents, preservatives, antioxidants, tonicity controlling agents, absorption delayingagents, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active substance, its use in the therapeutic compositions may be contemplated.

[0316] Illustrative, non-limiting carriers for use in formulating the pharmaceutical compositions include, for example, oil-in-water or water-in-oil emulsions, aqueous compositions with or without inclusion of organic co-solvents suitable for intravenous (IV) or intraocularly use, liposomes or surfactant-containing vesicles, microspheres, microbeads and microsomes, powders, tablets, capsules, suppositories, aqueous suspensions, aerosols, and other carriers apparent to one of ordinary skill in the art.

[0317] In particular embodiments, any antibody or antibody fragment, nucleic acid, the nucleic acid expression cassette or the vector herein described that is part of the pharmaceutical composition is provided in a liposome or lipid nanoparticle.

[0318] Pharmaceutical compositions as intended herein may be formulated for essentially any route of administration, such as without limitation, oral administration (such as, e.g., oral ingestion or inhalation), intranasal administration (such as, e.g., intranasal inhalation or intranasal mucosal application), parenteral administration (such as, e.g., subcutaneous, intravenous (I.V.), intraocularly, intramuscular, intraperitoneal, intrathecal or intracisternal injection or infusion), transdermal ortransmucosal (such as, e.g., oral, sublingual, intranasal) administration, topical administration, rectal, vaginal or intra-tracheal instillation, and the like. In this way, the therapeutic effects attainable by the methods and compositions can be, for example, systemic, local, tissue-specific, etc., depending on the specific needs of a given application.

[0319] For example, for oral administration, pharmaceutical compositions may be formulated in the form of pills, tablets, lacquered tablets, coated (e.g., sugar-coated) tablets, granules, hard and soft gelatin capsules, aqueous, alcoholic or oily solutions, syrups, emulsions or suspensions.

[0320] For example, for oral or nasal aerosol or inhalation administration, pharmaceutical compositions may be formulated for administration in the form of aerosols or sprays, for example, as solutions, suspensions or emulsions of the compound as taught herein or their physiologically tolerable salts in a pharmaceutically acceptable solvent, such as ethanol or water, or a mixture of such solvents. If required, theformulation can also additionally contain other pharmaceutical auxiliaries such as surfactants, emulsifiers and stabilizers as well as a propellant.

[0321] Examples of carriers for administration via mucosal surfaces depend upon the particular route, e.g., oral, sublingual, intranasal, etc. and are generally known in the art.

[0322] For example, for parenteral administration, pharmaceutical compositions may be advantageously formulated as solutions, suspensions or emulsions with suitable solvents, diluents, solubilisers or emulsifiers, etc. as generally known in the art. Injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents. The antibodies and pharmaceutically acceptable salts thereof of the invention can also be lyophilised and the lyophilisates obtained used, for example, for the production of injection or infusion preparations.

[0323] Where aqueous formulations are preferred, such may comprise one or more surfactants. For example, the composition can be in the form of a micellar dispersion comprising at least one suitable surfactant, e.g., a phospholipid surfactant. Various types of phospholipids are known in the art. Typically, a surfactant: active substance molar ratio in an aqueous formulation will be from about 10:1 to about 1:10, more typically from about 5:1 to about 1:5, however any effective amount of surfactant may be used in an aqueous formulation to best suit the specific objectives of interest.

[0324] In preferred embodiments, the pharmaceutical composition comprising the antibody or antibody fragment of the invention or nucleic acid encoding the antibody or antibody fragment, as taught herein is administered parenterally. More preferably, the pharmaceutical composition as taught herein is administered intravenously, for example by infusion, intrathecally, or intraocularly.

[0325] When rectally administered in the form of suppositories, these formulations may be prepared by mixing the compounds according to the invention with a suitable nonirritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquidity and / or dissolve in the rectal cavity to release the drug.

[0326] One skilled in this art will recognize that the above description is illustrative rather than exhaustive. Indeed, many additional formulations techniques andpharmaceutically acceptable excipients and carrier solutions are well-known to those skilled in the art, as is the development of suitable dosing and treatment regimens for using the particular compositions described herein in a variety of treatment regimens.

[0327] The present invention will be now described in more details, referring to examples that are not limitative.

[0328] Overview of sequences

[0329] SEQ ID N°:l (mature human Wnt7a) LGASIICNKIPGLAPRQRAICQSRPDAIIVIGEGSQMGLDECQFQFRNGRWNCSALGERTVFG KELKVGSREAAFTYAIIAAGVAHAITAACTQGNLSDCGCDKEKQGQYHRDEGWKWGGCSA DIRYGIGFAKVFVDAREIKQNARTLMNLHNNEAGRKILEENMKLECKCHGVSGSCTTKTCWT TLPQFRELGYVLKDKYNEAVHVEPVRASRNKRPTFLKIKKPLSYRKPMDTDLVYIEKSPNYCEE DPVTGSVGTQGRACNKTAPQASGCDLMCCGRGYNTHQYARVWQCNCKFHWCCYVKCNTC SERTEMYTCK

[0330] SEQ ID N°:2 (mature human Wnt7b) LGANIICNKIPGLAPRQRAICQSRPDAIIVIGEGAQMGINECQYQFRFGRWNCSALGEKTVFG QELRVGSREAAFTYAITAAGVAHAVTAACSQGNLSNCGCDREKQGYYNQAEGWKWGGCSA DVRYGIDFSRRFVDAREIKKNARRLMNLHNNEAGRKVLEDRMQLECKCHGVSGSCTTKTCW TTLPKFREVGHLLKEKYNAAVQVEVVRASRLRQPTFLRIKQLRSYQKPMETDLVYIEKSPNYCE EDAATGSVGTQGRLCNRTSPGADGCDTMCCGRGYNTHQYTKVWQCNCKFHWCCFVKCNT

[0331] SEQ ID N°:3 (human LRP5) MEAAPPGPPWPLLLLLLLLLALCGCPAPAAASPLLLFANRRDVRLVDAGGVKLESTIVVSGLED AAAVDFQFSKGAVYWTDVSEEAIKQTYLNQTGAAVQNVVISGLVSPDGLACDWVGKKLYWT DSETNRIEVANLNGTSRKVLFWQDLDQPRAIALDPAHGYMYWTDWGETPRIERAGMDGSTR KIIVDSDIYWPNGLTIDLEEQKLYWADAKLSFIHRANLDGSFRQKVVEGSLTHPFALTLSGDT LYWTDWQTRSIHACNKRTGGKRKEILSALYSPMDIQVLSQERQPFFHTRCEEDNGGCSHLCL LSPSEPFYTCACPTGVQLQDNGRTCKAGAEEVLLLARRTDLRRISLDTPDFTDIVLQVDDIRHA IAIDYDPLEGYVYWTDDEVRAIRRAYLDGSGAQTLVNTEINDPDGIAVDWVARNLYWTDTGT DRIEVTRLNGTSRKILVSEDLDEPRAIALHPVMGLMYWTDWGENPKIECANLDGQERRVLVN ASLGWPNGLALDLQEGKLYWGDAKTDKIEVINVDGTKRRTLLEDKLPHIFGFTLLGDFIYWTD WQRRSIERVHKVKASRDVIIDQLPDLMGLKAVNVAKVVGTNPCADRNGGCSHLCFFTPHAT RCGCPIGLELLSDMKTCIVPEAFLVFTSRAAIHRISLETNNNDVAIPLTGVKEASALDFDVSNN HIYWTDVSLKTISRAFMNGSSVEHVVEFGLDYPEGMAVDWMGKNLYWADTGTNRIEVARLD GQFRQVLVWRDLDNPRSLALDPTKGYIYWTEWGGKPRIVRAFMDGTNCMTLVDKVGRAND LTIDYADQRLYWTDLDTNMIESSNMLGQERVVIADDLPHPFGLTQYSDYIYWTDWNLHSIER ADKTSGRNRTLIQGHLDFVMDILVFHSSRQDGLNDCMHNNGQCGQLCLAIPGGHRCGCAS HYTLDPSSRNCSPPTTFLLFSQKSAISRMIPDDQHSPDLILPLHGLRNVKAIDYDPLDKFIYWV DGRQNIKRAKDDGTQPFVLTSLSQGQNPDRQPHDLSIDIYSRTLFWTCEATNTINVHRLSGE AMGVVLRGDRDKPRAIVVNAERGYLYFTNMQDRAAKIERAALDGTEREVLFTTGLIRPVALVVDNTLGKLFWVDADLKRIESCDLSGANRLTLEDANIVQPLGLTILGKHLYWIDRQQQMIERVE KTTGDKRTRIQGRVAHLTGIHAVEEVSLEEFSAHPCARDNGGCSHICIAKGDGTPRCSCPVH LVLLQNLLTCGEPPTCSPDQFACATGEIDCIPGAWRCDGFPECDDQSDEEGCPVCSAAQFPC ARGQCVDLRLRCDGEADCQDRSDEADCDAICLPNQFRCASGQCVLIKQQCDSFPDCIDGSD ELMCEITKPPSDDSPAHSSAIGPVIGIILSLFVMGGVYFVCQRVVCQRYAGANGPFPHEYVSG TPHVPLNFIAPGGSQHGPFTGIACGKSMMSSVSLMGGRGGVPLYDRNHVTGASSSSSSSTK ATLYPPILNPPPSPATDPSLYNMDMFYSSNIPATARPYRPYIIRGMAPPTTPCSTDVCDSDYSA SRWKASKYYLDLNSDSDPYPPPPTPHSQYLSAEDSCPPSPATERSYFHLFPPPPSPCTDSS

[0332] SEQ ID N°:4 (human LRP6) MGAVLRSLLACSFCVLLRAAPLLLYANRRDLRLVDATNGKENATIVVGGLEDAAAVDFVFSHG LIYWSDVSEEAIKRTEFNKTESVQNVVVSGLLSPDGLACDWLGEKLYWTDSETNRIEVSNLD GSLRKVLFWQELDQPRAIALDPSSGFMYWTDWGEVPKIERAGMDGSSRFIIINSEIYWPNGL TLDYEEQKLYWADAKLN FIH KSN LDGTN RQAVVKGSLPH PFALTLFEDI LYWTD WSTH SI LAC NKYTGEGLREIHSDIFSPMDIHAFSQQRQPNATNPCGIDNGGCSHLCLMSPVKPFYQCACPT GVKLLENGKTCKDGATELLLLARRTDLRRISLDTPDFTDIVLQLEDIRHAIAIDYDPVEGYIYWT DDEVRAIRRSFIDGSGSQFVVTAQIAHPDGIAVDWVARNLYWTDTGTDRIEVTRLNGTMRKI LISEDLEEPRAIVLDPMVGYMYWTDWGEIPKIERAALDGSDRVVLVNTSLGWPNGLALDYDE GKIYWGDAKTDKIEVMNTDGTGRRVLVEDKIPHIFGFTLLGDYVYWTDWQRRSIERVHKRS AEREVIIDQLPDLMGLKATNVHRVIGSNPCAEENGGCSHLCLYRPQGLRCACPIGFELISDMK TCIVPEAFLLFSRRADIRRISLETNNNNVAIPLTGVKEASALDFDVTDNRIYWTDISLKTISRAF MNGSALEHVVEFGLDYPEGMAVDWLGKNLYWADTGTNRIEVSKLDGQHRQVLVWKDLDSP RALALDPAEGFMYWTEWGGKPKIDRAAMDGSERTTLVPNVGRANGLTIDYAKRRLYWTDLD TNLIESSNMLGLNREVIADDLPHPFGLTQYQDYIYWTDWSRRSIERANKTSGQNRTIIQGHL DYVMDILVFHSSRQSGWNECASSNGHCSHLCLAVPVGGFVCGCPAHYSLNADNRTCSAPTT FLLFSQKSAINRMVIDEQQSPDIILPIHSLRNVRAIDYDPLDKQLYWIDSRQNMIRKAQEDGS QGFTVVVSSVPSQNLEIQPYDLSIDIYSRYIYWTCEATNVINVTRLDGRSVGVVLKGEQDRPR AVVVNPEKGYMYFTNLQERSPKIERAALDGTEREVLFFSGLSKPIALALDSRLGKLFWADSDL RRIESSDLSGANRIVLEDSNILQPVGLTVFENWLYWIDKQQQMIEKIDMTGREGRTKVQARI AQLSDIHAVKELNLQEYRQHPCAQDNGGCSHICLVKGDGTTRCSCPMHLVLLQDELSCGES QFQCASGQCIDGALRCNGDANCQDKSDEKNCEVLCLIDQFRCANGQCIGKHKKCDHNVDC SDKSDELDCYPTEEPAPQATNTVGSVIGVIVTIFVSGTVYFICQRMLCPRMKGDGETMTNDYV VHGPASVPLGYVPHPSSLSGSLPGMSRGKSMISSLSIMGGSSGPPYDRAHVTGASSSSSSST KGTYFPAILNPPPSPATERSHYTMEFGYSSNSPSTHRSYSYRPYSYRHFAPPTTPCSTDVCDSD YAPSRRMTSVATAKGYTSDLNYDSEPVPPPPTPRSQYLSAEENYESCPPSPYTERSYSHHLYPP PPSPCTDSS

[0333] SEQ ID N°:5 (human LRP6) MGAVLRSLLACSFCVLLRAAPLLLYANRRDLRLVDATNGKENATIVVGGLEDAAAVDFVFSHG LIYWSDVSEEAIKRTEFNKTESVQNVVVSGLLSPDGLACDWLGEKLYWTDSETNRIEVSNLD GSLRKVLFWQELDQPRAIALDPSSGFMYWTDWGEVPKIERAGMDGSSRFIIINSEIYWPNGL TLDYEEQKLYWADAKLN FIH KSN LDGTN RQAVVKGSLPH PFALTLFEDI LYWTD WSTH SI LAC NKYTGEGLREIHSDIFSPMDIHAFSQQRQPNATNPCGIDNGGCSHLCLMSPVKPFYQCACPT GVKLLENGKTCKDGATELLLLARRTDLRRISLDTPDFTDIVLQLEDIRHAIAIDYDPVEGYIYWT DDEVRAIRRSFIDGSGSQFVVTAQIAHPDGIAVDWVARNLYWTDTGTDRIEVTRLNGTMRKI LISEDLEEPRAIVLDPMVGYMYWTDWGEIPKIERAALDGSDRVVLVNTSLGWPNGLALDYDE GKIYWGDAKTDKIEVMNTDGTGRRVLVEDKIPHIFGFTLLGDYVYWTDWQRRSIERVHKRS AEREVIIDQLPDLMGLKATNVHRVIGSNPCAEENGGCSHLCLYRPQGLRCACPIGFELISDMKTCIVPEAFLLFSRRADIRRISLETNNNNVAIPLTGVKEASALDFDVTDNRIYWTDISLKTISRAF MNGSALEHVVEFGLDYPEGMAVDWLGKNLYWADTGTNRIEVSKLDGQHRQVLVWKDLDSP RALALDPAEGFMYWTEWGGKPKIDRAAMDGSERTTLVPNVGRANGLTIDYAKRRLYWTDLD TNLIESSNMLGLNREVIADDLPHPFGLTQYQDYIYWTDWSRRSIERANKTSGQNRTIIQGHL DYVMDILVFHSSRQSGWNECASSNGHCSHLCLAVPVGGFVCGCPAHYSLNADNRTCSAPTT FLLFSQKSAINRMVIDEQQSPDIILPIHSLRNVRAIDYDPLDKQLYWIDSRQNMIRKAQEDGS QGFTVVVSSVPSQNLEIQPYDLSIDIYSRYIYWTCEATNVINVTRLDGRSVGVVLKGEQDRPR AIVVNPEKGYMYFTNLQERSPKIERAALDGTEREVLFFSGLSKPIALALDSRLGKLFWADSDLR RIESSDLSGANRIVLEDSNILQPVGLTVFENWLYWIDKQQQMIEKIDMTGREGRTKVQARIA QLSDIHAVKELNLQEYRQHPCAQDNGGCSHICLVKGDGTTRCSCPMHLVLLQDELSCGESQ FQCASGQCIDGALRCNGDANCQDKSDEKNCEVLCLIDQFRCANGQCIGKHKKCDHNVDCS DKSDELDCYPTEEPAPQATNTVGSVIGVIVTIFVSGTVYFICQRMLCPRMKGDGETMTNDYVV HGPASVPLGYVPHPSSLSGSLPGMSRGKSMISSLSIMGGSSGPPYDRAHVTGASSSSSSSTK GTYFPAILNPPPSPATERSHYTMEFGYSSNSPSTHRSYSYRPYSYRHFAPPTTPCSTDVCDSDY APSRRMTSVATAKGYTSDLNYDSEPVPPPPTPRSQYLSAEENYESCPPSPYTERSYSHHLYPPP PSPCTDSS

[0334] SEQ ID N°:6 (human Reck) MATVRASLRGALLLLLAVAGVAEVAGGLAPGSAGALCCNHSKDNQMCRDVCEQIFSSKSES RLKHLLQRAPDYCPETMVEIWNCMNSSLPGVFKKSDGWVGLGCCELAIALECRQACKQASS KNDISKVCRKEYENALFSCISRNEMGSVCCSYAGHHTNCREYCQAIFRTDSSPGPSQIKAVE NYCASISPQLIHCVNNYTQSYPMRNPTDSLYCCDRAEDHACQNACKRILMSKKTEMEIVDGLI EGCKTQPLPQDPLWQCFLESSQSVHPGVTVHPPPSTGLDGAKLHCCSKANTSTCRELCTKLY SMSWGNTQSWQEFDRFCEYNPVEVSMLTCLADVREPCQLGCRNLTYCTNFNNRPTELFRSC NAQSDQGAMNDMKLWEKGSIKMPFINIPVLDIKKCQPEMWKAIACSLQIKPCHSKSRGSIIC KSDCVEILKKCGDQNKFPEDHTAESICELLSPTDDLKNCIPLDTYLRPSTLGNIVEEVTHPCNP NPCPANELCEVNRKGCPSGDPCLPYFCVQGCKLGEASDFIVRQGTLIQVPSSAGEVGCYKICS CGQSGLLENCMEMHCIDLQKSCIVGGKRKSHGTSFSIDCNVCSCFAGNLVCSTRLCLSEHSS EDDRRTFTGLPCNCADQFVPVCGQNGRTYPSACIARCVGLQDHQFEFGSCMSKDPCNPNPC QKNQRCIPKPQVCLTTFDKFGCSQYECVPRQLACDQVQDPVCDTDHMEHNNLCTLYQRGKS LSYKGPCQPFCRATEPVCGHNGETYSSVCAAYSDRVAVDYYGDCQAVGVLSEHSSVAECAS VKCPSLLAAGCKPIIPPGACCPLCAGMLRVLFDKEKLDTIAKVTNKKPITVLEILQKIRMHVSVP QCDVFGYFSIESEIVILIIPVDHYPKALQIEACNKEAEKIESLINSDSPTLASHVPLSALIISQVQ VSSSVPSAGVRARPSCHSLLLPLSLGLALHLLWTYN

[0335] SEQ ID N°:7 (human Gprl24) / Adhesion G protein-coupled receptor A2 (ADGRA2) MGAGGRRMRGAPARLLLPLLPWLLLLLAPEARGAPGCPLSIRSCKCSGERPKGLSGGVPGPA RRRVVCSGGDLPEPPEPGLLPNGTVTLLLSNNKITGLRNGSFLGLSLLEKLDLRNNIISTVQPG AFLGLGELKRLDLSNNRIGCLTSETFQGLPRLLRLNISGNIFSSLQPGVFDELPALKVVDLGTEF LTCDCHLRWLLPWAQNRSLQLSEHTLCAYPSALHAQALGSLQEAQLCCEGALELHTHHLIPSL RQVVFQGDRLPFQCSASYLGNDTRIRWYHNRAPVEGDEQAGILLAESLIHDCTFITSELTLSH IGVWASGEWECTVSMAQGNASKKVEIVVLETSASYCPAERVANNRGDFRWPRTLAGITAYQ SCLQYPFTSVPLGGGAPGTRASRRCDRAGRWEPGDYSHCLYTNDITRVLYTFVLMPINASNA LTLAHQLRVYTAEAASFSDMMDVVYVAQMIQKFLGYVDQIKELVEVMVDMASNLMLVDEHLL WLAQREDKACSRIVGALERIGGAALSPHAQHISVNARNVALEAYLIKPHSYVGLTCTAFQRRE GGVPGTRPGSPGQNPPPEPEPPADQQLRFRCTTGRPNVSLSSFHIKNSVALASIQLPPSLFSSLPAALAPPVPPDCTLQLLVFRNGRLFHSHSNTSRPGAAGPGKRRGVATPVIFAGTSGCGVGNL TEPVAVSLRHWAEGAEPVAAWWSQEGPGEAGGWTSEGCQLRSSQPNVSALHCQHLGNVA VLMELSAFPREVGGAGAGLHPVVYPCTALLLLCLFATIITYILNHSSIRVSRKGWHMLLNLCFHI AMTSAVFAGGITLTNYQMVCQAVGITLHYSSLSTLLWMGVKARVLHKELTWRAPPPQEGDPA LPTPSPMLRFYLIAGGIPLIICGITAAVNIHNYRDHSPYCWLVWRPSLGAFYIPVALILLITWIYF LCAGLRLRGPLAQNPKAGNSRASLEAGEELRGSTRLRGSGPLLSDSGSLLATGSARVGTPGP PEDGDSLYSPGVQLGALVTTHFLYLAMWACGALAVSQRWLPRVVCSCLYGVAASALGLFVFT HHCARRRDVRASWRACCPPASPAAPHAPPRALPAAAEDGSPVFGEGPPSLKSSPSGSSGHPL ALGPCKLTNLQLAQSQVCEAGAAAGGEGEPEPAGTRGNLAHRHPNNVHHGRRAHKSRAKG HRAGEACGKNRLKALRGGAAGALELLSSESGSLHNSPTDSYLGSSRNSPGAGLQLEGEPMLT PSEGSDTSAAPLSEAGRAGQRRSASRDSLKGGGALEKESHRRSYPLNAASLNGAPKGGKYD DVTLMGAEVASGGCMKTGLWKSETTV

[0336] SEQ ID N°:9 (human Fzdl) MAEEEAPKKSRAAGGGASWELCAGALSARLAEEGSGDAGGRRRPPVDPRRLARQLLLLLWL LEAPLLLGVRAQAAGQGPGQGPGPGQQPPPPPQQQQSGQQYNGERGISVPDHGYCQPISIP LCTDIAYNQTIMPNLLGHTNQEDAGLEVHQFYPLVKVQCSAELKFFLCSMYAPVCTVLEQALP PCRSLCERARQGCEALMNKFGFQWPDTLKCEKFPVHGAGELCVGQNTSDKGTPTPSLLPEF WTSNPQHGGGGHRGGFPGGAGASERGKFSCPRALKVPSYLNYHFLGEKDCGAPCEPTKVYG LMYFGPEELRFSRTWIGIWSVLCCASTLFTVLTYLVDMRRFSYPERPIIFLSGCYTAVAVAYIAG FLLEDRVVCNDKFAEDGARTVAQGTKKEGCTILFMMLYFFSMASSIWWVILSLTWFLAAGMK WGHEAIEANSQYFHLAAWAVPAIKTITILALGQVDGDVLSGVCFVGLNNVDALRGFVLAPLFV YLFIGTSFLLAGFVSLFRIRTIMKHDGTKTEKLEKLMVRIGVFSVLYTVPATIVIACYFYEQAFRD QWERSWVAQSCKSYAIPCPHLQAGGGAPPHPPMSPDFTVFMIKYLMTLIVGITSGFWIWSG KTLNSWRKFYTRLTNSKQGETTV

[0337] SEQ ID N°: 45 (hADAMTSlO (823-1103)) SLPPYSWHYAPWTKCSAQCAGGSQVQAVECRNQLDSSAVAPHYCSAHSKLPKRQRACNTE PCPPDWVVGNWSLCSRSCDAGVRSRSVVCQRRVSAAEEKALDDSACPQPRPPVLEACHGPT CPPEWAALDWSECTPSCGPGLRHRVVLCKSADHRATLPPAHCSPAAKPPATMRCNLRRCPPA RWVAGEWGECSAQCGVGQRQRSVRCTSHTGQASHECTEALRPPTTQQCEAKCDSPTPGD GPEECKDVNKVAYCPLVLKFQFCSRAYFRQMCCKTCHGH

[0338] SEQ ID N°: 46 (hWnt7a linker (171-199)) VEPVRASRNKRPTFLKIKKPLSYRKPMDT

[0339] SEQ ID N°: 47 (hReck CK1-5 (27-340)) GLAPGSAGALCCNHSKDNQMCRDVCEQIFSSKSESRLKHLLQRAPDYCPETMVEIWNCMNS SLPGVFKKSDGWVGLGCCELAIALECRQACKQASSKNDISKVCRKEYENALFSCISRNEMGS VCCSYAGHHTNCREYCQAIFRTDSSPGPSQIKAVENYCASISPQLIHCVNNYTQSYPMRNPT DSLYCCDRAEDHACQNACKRILMSKKTEMEIVDGLIEGCKTQPLPQDPLWQCFLESSQSVHP GVTVHPPPSTGLDGAKLHCCSKANTSTCRELCTKLYSMSWGNTQSWQEFDRFCEYNPVEVS MLTCLA

[0340] SEQ ID N°: 48 (Wnt7b linker)

[0341] VEVVRASRLRQPTFLRLKQLRSYQKPMETSEQ ID N°: 49 (CK4 motif RECK) CCDRAEDHACQNACKRILMSKKTEMEIVDGLIEGCKTQPLPQDPLWQC

[0342] SEQ ID N°: 50 (CK5 motif RECK)

[0343] CCS KANTSTC RE LCTKLYS M S WG NTQS WQE FD RFCEYN PVE VS M LTC

[0344] SEQ ID N°: 51 (RECK binding domain) HVEPVRASRNKRPTFLKIKKPLSYRKPMDTDLVYIEKSPNYC

[0345] SEQ ID N°: 52 (RECK binding domain) YEWRASRLRQPTFLRIKQLRSYQKPMET

[0346] SEQ ID N°: 54 (RECK BINDING DOMAIN) XXXVXAXRXXXXXFLXIXXXXXYXKXXXX

[0347] SEQ ID N°: 55 (RECK binding domain)

[0348] VXAXRXXXXXFLXIXXXXXYXK

[0349] SEQ ID N°: 56 (RECK binding domain) XXXVXAXRXXXXXFLXXXXXXXXXKXXXX

[0350] SEQ ID N°: 57 (RECK binding domain)

[0351] VXAXRXXXXXFLXXXXXXXXXK

[0352] SEQ ID N°: 58 (N-TERMINAL DOMAIN GPR124) APGCPLSIRSCKCSGERPKGLSGGVPGPARRRVVCSGGDLPEPPEPGLLPNGTVTLLLSNNKI TGLRNGSFLGLSLLEKLDLRNNIISTVQPGAFLGLGELKRLDLSNNRIGCLTSETFQGLPRLLRL NISGNIFSSLQPGVFDELPALKVVDLGTEFLTCDCHLRWLLPWAQNRSLQLSEHTLCAYPSAL HAQALGSLQEAQLCCEGALELHTHHLIPSLRQVVFQGDRLPFQCSASYLGNDTRIRWYHNRA PVEGDEQAGILLAESLIHDCTFITSELTLSHIGVWASGEWECTVSMAQGNASKKVEIVVLETS ASYCPAERVANNRGDFRWPRTLAGITAYQSCLQYPFTSVPLGGGAPGTRASRRCDRAGRWE PGDYSHCLYTNDITRVLYTFVLMPINASNALTLAHQLRVYTAEAASFSDMMDVVYVAQMIQKF LGYVDQIKELVEVMVDMASNLMLVDEHLLWLAQREDKACSRIVGALERIGGAALSPHAQHIS VNARNVALEAYLIKPHSYVGLTCTAFQRREGGVPGTRPGSPGQNPPPEPEPPADQQLRFRCTT GRPNVSLSSFHIKNSVALASIQLPPSLFSSLPAALAPPVPPDCTLQLLVFRNGRLFHSHSNTSR PG AAG PG KRRG VATPVI FAGTSGCG VG N LTE PVAVSLRH WAEG AE PVAAW WSQEG PG E AG GWTSEGCQLRSSQPNVSALHCQHLGNVAVLMELSAFPREVGGAGAGLHP

[0353] Anti-Gprl24 antibody fragmentCD CD CD

[0354] FR FR FR FR CD CD CD FRL FRL FRL FRL ID VH RH RH RH VL

[0355] Hl H2 H3 H4 RL1 RL2 RL3 1 2 3 4 1 2 3

[0356] A10 GV PD RLT QAVLTQP RFS QV ISV PSASGTP GS QVQLQESG QL DTS GQRVTIS KS PGLVKPSET QE RS CSGSSS QA GT LSLTCTVSG SG QF NIGSNYV VLT SAS GSISGSSYY PG SLK YWYQQL QP WY LAI WSWIRQPP NIY LVK Wl LSS PGTAPKL AA PSA QQ SG GRGLEWIG YS PSE RQ VTA WG LIYRNNQ SG WD SG LP LRS NIYYSGSTY GS LLV TLS PP AD QG RPSGVPD SSS RN DS TP GT ED FG YNPSLKSRL TYY QG LTC GR TAV TM RFSGSKS NIG NQ LS GQ AP EA GG TISVDTSRS GS NP VD TVS GL YY VTV GTSASLAI SN RP GV RVT KLL DY TKL QFSLKLSSV SYY SLK AF GG EWI CA SS SGLRSED YVY S V ISC IY YC TVL TAADTAVYY WS S DI SIS G R (SE EADYYCA (SE (SE (SE (SE (SE (SE (SE CARLLVQG (SE (SE (SE (SE (SE (SE Q AWDDSL Q Q Q Q Q Q Q VDAFDIWG Q Q Q Q Q Q ID SGWFG ID ID ID ID ID ID ID QGTMVTVS ID ID ID ID ID ID N°: GGTKLTV N°: N°: N°: N°: N°: N°: N°: S (SEQ ID N°: N°: N°: N°: N°: N°: 102 L (SEQ ID 103 105 107 110 113 115 117 N°: 63) 89) 91) 93) 95) 97) 99) ) N°: 64) ) ) ) ) ) ) ) C03 GV PD RLT RFS ISV SYVLTQP GS QV DT PSASGTP KS QVQLQESG QL SR GQRVTIS GS PGLVKPSET QE NQ CSGSSS SYV SAS LSLTCTVSG SG FSL NIGSNYV LTQ LAI GSISGSSYY PG KLS YWYQQL PPS WY SG WSWIRQPP NIY LVK Wl SVT PGTAPKL AA AS QQ LQ GKGLEWIG YS PSE RQ AA WG LIYRNNQ SG WD GT LP SE NIYYSGSTY GS LLV TLS PP DT QG RPSGVPD SSS RN DS PG GT DE FG YNPSLKSRL TYY QG LTC GK AVY TM RFSGSKS NIG NQ LN QR AP AD GG TISVDTSRN GS NP VD TVS GL YC VTV GSSASLA SN RP GV VTI KLL YY TKL QFSLKLSSV SYY SLK AF GG EWI AR SS ISGLQSE YVY S V SC IY C TVL TAADTAVYY WS S DI SIS G (SE (SE DEADYYC (SE (SE (SE (SE (SE (SE (SE CARLLVQG (SE (SE (SE (SE (SE Q Q AAWDDS Q Q Q Q Q Q Q VDAFDIWG Q Q Q Q Q ID ID LNGWFG ID ID ID ID ID ID ID QGTMVTVS ID ID ID ID ID N°: N°: GGTKLTV N°: N°: N°: N°: N°: N°: N°: S (SEQ ID N°: N°: N°: N°: N°: 100 102 L (SEQ ID 103 105 108 111 113 116 117 N°: 65) 89) 91) 93) 95) 98) ) ) N°: 66) ) ) ) ) ) ) ) A05 QV RVT SYELTQP SYE GV QVQLQESG QL MS PSASGTP LTQ PD PGLVKPSET QE VD GQRVTIS PPS WY RFS LSLSCTVSG TIY SG Wl TSK CSGSSS AT AS QQ GS DSISSRSHY YS PG RQ NQ WG NIGTNYV SG WD GT LP KS WGWIRQP GN LR LVK PP FSL QG YWYQQL SSS SD DS PG GM GT FG PGKGLEWI SR TYY GS PSE GK NL TM PGMAPKL NIG DQ LS QR AP SAS GG GTIYYSGNT SH NP VD TLS GL SSV VTV LIYSDDQ TNY RP GV VTI KLL LAI TKL YYNPSLKSR YW SLK AF LS EWI TAA SS RPSGVPD VY S V SC IY SG TVL VTMSVDTS G S DI CT G DT (SE RFSGSKS (SE (SE (SE (SE (SE LRS (SE KNQFSLNL (SE (SE (SE VS (SE AVY Q GTSASLAI Q Q Q Q Q ED Q SSVTAADTA Q Q Q GD Q FC ID SGLRSED ID ID ID ID ID EA ID VYFCAILRG ID ID ID SIS ID Al N°: EADYYCA N°: N°: N°: N°: N°: DY N°: SVDAFDIW N°: N°: N°: (SE N°: (SE 102 TWDDSL 104 106 109 112 114 YC 117

[0357]

[0358] GQGTMVTV 90) 92) 94) Q 98) Q ) SGWFG ) ) ) ) ) (SE )SS (SEQ ID ID ID GGTKLTV Q N°: 67) N°: N°: L (SEQ ID ID 96) 101 N°: 68) N°:

[0359] ) 115

[0360]

[0361] ) Anti-Reck antibody fragment

[0362] CD CD CD FR FR FR FR CD CD CD FRL FRL FRL FRL ID VH RH RH RH VL

[0363] Hl H2 H3 H4 RL1 RL2 RL3 1 2 3 4 1 2 3

[0364] 01A08 RLT GV QL MS PS QL VD RFS QLQLQESG QE TSK DIVMTQS GS PGRVKPSE SG NQ PSSLSAS GS TLSLTCKVS PG FSL VGDRVTI DIV GT GFSISRGYY RV KM TCRASQS MT DFT WAWVRQA KPS WV RS ISRYLNW QS WY LTI PGKGLEWI NIH AFY ETL RQ VTA YQQKPG PSS QQ SSL GNIHHSGS HS GS SLT AP AD WG KAPKLLM RA LSA KP QP TYYSPSLKS GS GN CK GK TAV QG YAASSLQ SQ QQ SV GK ED FG RLTMSVDT RG TYY YYV VS GL YF TTV SGVPSRF SIS AA SYS GD AP FAT GG SKNQFSLK YY SPS TAL GF EWI CA IVS SGSGSGT RYL SSL TPL RVT KLL YY TKV MRSVTAAD WA LKS DV SIS G R S DFTLTISS N QS T ITC MY C EIK TAVYFCARA (SE (SE (SE (SE (SE (SE (SE LQPEDFA (SE (SE (SE (SE (SE (SE (SE FYGSGNYY Q Q Q Q Q Q Q TYYCQQS Q Q Q Q Q Q Q VTALDVWG ID ID ID ID ID ID ID YSTPLTF ID ID ID ID ID ID ID QGTTVIVSS N°: N°: N°: N°: N°: N°: N°: GGGTKVE N°: N°: N°: N°: N°: N°: N°: (SEQ ID N0: 118 122 126 130 134 138 142 IK (SEQ 146 150 154 158 162 165 168 69) ) ) ) ) ) ) ) ID N°: 70) ) ) ) ) ) ) ) 01F11 RFT GV EV ISR PS QL DN RFS EVQLVESG VES SN DIQLTQS GS GALVQPGG GG NR PSSLSAS GS SLRLSCAAS ALV LYL VGDRVTI DIQ GT GFTVSSNY AEL QP QM TCRASQ LTQ DFT MSWVRQA VIY LPF GG WV NS GISSYLA SPS WY LTI PGKGLEWV SG RV SLR RQ LRA WYQQKP SLS QQ SG SVIYSGGST GS GV LS AP EDT WG GKAPKLLI RA AS KP LQ YYAGSVKG TYY VIS CA GK GV QG YAASTLQ SQ QQ VG GK AE FG RFTISRDNS SN AG KYY AS GL YY TTV SGVPSRF GIS AA SYS DR AP DF QG NNRLYLQM YM SVK GL GFT EW CA TVS SGSGSGT SYL STL SPI VTI KLL ATY TRL NSLRAEDT S G DV VS VS R S DFTLTISG A QS T TC IY YC DIK GVYYCARA (SE (SE (SE (SE (SE (SE (SE LQAEDFA (SE (SE (SE (SE (SE (SE (SE ELLPFRVGV Q Q Q Q Q Q Q TYYCQQS Q Q Q Q Q Q Q VISKYYGLD ID ID ID ID ID ID ID YSSPITFG ID ID ID ID ID ID ID VWGQGTTV N°: N°: N°: N°: N°: N°: N°: QGTRLDI N°: N°: N°: N°: N°: N°: N°: TVSS (SEQ 119 123 127 131 135 139 143 K (SEQ ID 147 151 155 159 163 166 169 ID N°: 71) ) ) ) ) ) ) ) N°: 72) ) ) ) ) ) ) ) 03A04 EVQLVESG SIIA NS EV WV RFT WG DIQMTQS RA AA DIQ WY GV FG GGLVQPG NY SD GR QL RQ ISR QG PSSLSAS SQ SN HQ MT QQ PS QG GSLRLACTT AM VST TIF VES TP DN TLV AGDRVTI SIN LQ TYT QS KP RFS TKL SGFTFSNY S YYA GV GG GG SN TVS TCRASQS TYL S THT PSS GK GS EIK AMSWVRQ (SE DS VMI GL RLE DTL S INTYLNW N (SE (SE LSA AP GS (SE TPGGRLEW Q VR GE VQ WV YL (SE YQQKPG (SE Q Q SA KLL GT Q VSSIIASDV ID G YG PG S QM Q KAPKLLIY Q ID ID GD IY DFT ID

[0365]

[0366] STYYADSVR N°: (SE FD GS (SE HS ID AASNLQS ID N°: N°: RVT (SE LTI N°:GRFTISRDN 120 Q S LRL Q LR N°: GVPSRFS N°: 152 156 ITC Q SSL 170 SNDTLYLQ ) ID (SE AC ID DE 144 GSGSGT 148 ) ) (SE ID QP ) MHSLRDED N°: Q TTS N°: DT ) DFTLTISS ) Q N°: ED TAVYFCAK 124 ID GFT 136 AVY LQPEDFA ID 163 FAT NSGRTIFGV ) N°: FS ) FC TYYCHQT N°: ) YY VMIGEYGF 128 (SE AK YTTHTFG 160 C DSWGQGT ) Q (SE QGTKLEI ) (SE LVTVSS ID Q K (SEQ ID Q (SEQ ID N0: N°: ID N°: 74) ID 73) 132 N°: N°:

[0367] ) 140 165 ) ) 07B05 RVT GIP QV ISV QSVLTQP DR QVQLVESG QL DM PSVSAAP FTG PGLVKPSQ VES SK GQHVTIS QS SKS TLSLTCTVS GP NQ CSGSSS VLT GT GGSISSGD GL FSL NVGKNF QP SAT YYWSWIRQ YIY VKP Wl KLS VSWYQQ PSV WY LGI HPGKGLE YS SQ RQ SVT LPGTVPK SG SA QQ TGL WIGYIYYSG GS ERL TLS HP AA WG LLIYDNN SSS GT AP LP QT STYHNPSL SG TYH KG LTC GK DT QG RRPSGIP NV DN WD GQ GT GD FG KSRVTISVD DY NP LD TVS GL AVY TM DRFTGSK GK NR TSL HV VPK EA GG MSKNQFSL YW SLK AF GG EWI YC VTV SGTSATL NF RP SA TIS LLI DY TRL KLSSVTAAD S S DI SIS G AR SS GITGLQT VS S W C Y YC TVL TAVYYCARE (SE (SE (SE (SE (SE (SE (SE GDEADYY (SE (SE (SE (SE (SE (SE (SE RLKGLDAF Q Q Q Q Q Q Q CGTWDT Q Q Q Q Q Q Q DIWGQGT ID ID ID ID ID ID ID SLSAWF ID ID ID ID ID ID ID MVTVSS N°: N°: N°: N°: N°: N°: N°: GGGTRLT N°: N°: N°: N°: N°: N°: N°: (SEQ ID N0: 121 125 129 133 137 141 145 VL (SEQ 149 153 157 161 164 167 171

[0368]

[0369] 75) ) ) ) ) ) ) ) ID N°: 76) ) ) ) ) ) ) )

[0370] SEQ ID N°: 172 (5-mer linker)

[0371] GSGGS

[0372] Anti-Fzd sequences

[0373] ID VH CDR1 CDR CDR3 VL CDR1 CDR CDR3

[0374] 2 2

[0375] 1 EVQLVESGGGLVKPGG FTFTN SAIS CARAT DIQMTQSPSSLSASVGDRVTI RASQ AAS CQQSY SLRLSCAASGFTFTNYA YAMS GSG GFGTV TCRASQSISSYLNWYQQKPG SISSY SLQ STPLTF MSWVRQAPGKGLEWV (SEQ GST VFDY KAPKLLIYAASSLQSGVPSRFS LN S (SEQ SAISGSGGSTYYADSVK ID N°: YYA W GSGSGTDFTLTISSLQPEDFAT (SEQ (SE ID N°: GRFTISRDDSKNTLYLQ 180) (SE (SEQ YYCQQSYSTPLTFGGGTKVEI ID N°: Q ID 191) MNSLKTEDTAVYYCAR Q ID ID N°: KR (SEQ ID N°: 177) 187) N°:

[0376] ATGFGTWFDYWGQG N°: 185) 189) TLVTVSS (SEQ ID N0: 182)

[0377] 174)

[0378] 2 EVQLVESGGGLVQPG NSSFY TVY AFPGS DIQMTQSPSSLSASVGDRVTI SVSSA SAS SSYSLI GSLRLSCAASGFNSSF FM PYL YHPM TCRASQSVSSAVAWYQQKPG (SEQ SLY (SEQ YFMHWVRQAPGKGLE (SEQ DYT (SEQ KAPKLLIYSASSLYSGVPSRFS ID N°: S ID N°: WVATVYPYLDYTYYAD ID N°: Y ID N°: GSRSGTDFTLTISSLQPEDFAT 188) (SE 192) SVKGRFTISADTSKNTA 181) (SE 186) YYCQQSSYSLITFGQGTKVEIK Q ID YLQMNSLRAEDTAVYY Q ID LEDKTHTKV (SEQ ID N0: 178) N°:

[0379] CARAFPGSYHPMDYW N°: 190) GQGTLVTVSS (SEQ ID 183)

[0380] N°: 175)

[0381]

[0382] EVQLVESGGGLVQPG NSSFY ATV AFPGS DIQMTQSPSSLSASVGDRVTI SVSSA SAS VSSA GSLRLSCAASGFNSSF FM YPY YHPM TCRASQSVSSAVAWYQQKPG (SEQ SLY (SEQ YFMHWVRQAPGKGLE (SEQ LDY (SEQ KAPKLLIYSASSLYSGVPSRFS ID N°: S ID N°: WVATVYPYLDYTYYAD ID N°: TY ID N°: GSRSGTDFTLTISSLQPEDFAT 188) (SE 193) SVKGRFTISADTSKNTA 181) (SE 186) YYCQQVSSATFGQGTKV Q ID YLQMNSLRAEDTAVYY Q ID (SEQ ID N°: 179) N°:

[0383] CARAFPGSYHPMDYW N°: 190) GQGTLVTVSS (SEQ ID 184)

[0384]

[0385] N°: 176)

Claims

CLAIMS1. A soluble antibody or antibody fragment, wherein the antibody or antibody fragment is a modulator of Wnt7 signaling pathway, preferably an agonist of the Wnt7 pathway, and wherein said antibody or antibody fragment comprises one or more Fzd binding domains, and further one or more Gprl24 and / or Reck binding domains.

2. The antibody or antibody fragment according to any one of the previous claims, wherein the one or more Fzd binding domains bind an amino acid sequence having at least 75% sequence identity to a fragment of the Fzd proteins as depicted in SEQ ID N°: 9.

3. The antibody or antibody fragment according to any one of the previous claims, wherein the one or more Gprl24 or Reck binding domains bind an amino acid sequence having at least 75% sequence identity to a fragment of the Gprl24 or Reck proteins as depicted in SEQ ID N°: 7 or 6.

4. The antibody or antibody fragment according to any of the previous claims, wherein at least one of the one or more Fzd binding domains and / or the one or more Gprl24 or Reck binding domains is derived from an antibody, an antibody fragment, an immunoglobulin single variable domain (ISVD) or VHH.

5. The antibody or antibody fragment according to any one of the previous claims, wherein said antibody or antibody fragment comprises an anti-Gprl24 IgG or scFv comprising a VH sequence according to SEQ ID N° 63, 65, or 67, and a VL sequence according to SEQ ID N° 64, 66, or 68, optionally separated by a linker sequence; preferably a VH sequence according to SEQ ID N° 63 and a VL sequence according to SEQ ID N° 64, VH sequence according to SEQ ID N°65 and a VL sequence according to SEQ ID N° 66, or a VH sequence according to SEQ ID N° 67 and a VL sequence according to SEQ ID N° 68.

6. The antibody or antibody fragment according to one of the previous claims, wherein said antibody or antibody fragment comprises an anti-Reck IgG or scFv comprising a VH sequence according to SEQ ID N° 69, 71, 73, or 75, and a VL sequence according to SEQ ID N° 70, 72, 74, or 76, optionally separated by a linker sequence; preferably comprising a VH sequence according to SEQ ID N° 69 and a VL sequence according to SEQ ID N° 70, a VH sequence according to SEQ ID N° 71 and a VL sequence according to SEQ ID 72, a VH sequence according to SEQ ID N° 73 and a VL sequence according to SEQ ID N° 74, or a VH sequence according to SEQ ID N° 75 and a VL sequence according to SEQ ID N° 76.

7. The antibody or antibody fragment according to any one of the previous claims, wherein said antibody or antibody fragment comprises an anti-Fzd IgG or scFv comprising a VH sequence according to SEQ ID N° 174, 175, or 176, and a VL sequence according to SEQ ID N° 177, 178, or 179, optionally separated by a linker sequence.

8. The antibody or antibody fragment according to any one of the previous claims, wherein the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains, and one or more Gprl24 and / or Reck binding domains.

9. The antibody or antibody fragment according to any one or the previous claims, wherein said antibody or antibody fragment is bispecific.

10. The antibody or antibody fragment according to claim 9, wherein the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains and one or more Gprl24 binding domains.

11. The antibody or antibody fragment according to claim 10, wherein said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains and one or more Gprl24 binding domains.

12. The antibody or antibody fragment according to claim 9, wherein the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains and one or more Reck binding domains.

13. The antibody or antibody fragment according to claim 12, wherein said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains and one or more Reck binding domains.

14. The antibody or antibody fragment according to claim 9, wherein said antibody or antibody fragment is a bispecific antibody with two binding domains, said binding domains including one Fzd binding domain, and one Gprl24 binding domain or Reck binding domain.

15. The antibody or antibody fragment according to any one of the previous claims 1-8, wherein said antibody or antibody fragment is trispecific and / or trivalent.

16. The antibody or antibody fragment according to any one of the previous claims 1-8, or 15, comprising one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains.

17. The antibody or antibody fragment according to claim 15 or 16, comprising: i) one region that specifically binds to a set of one or more Fzd epitopes; ii) one region that specifically binds to a set of one or more Gprl24 epitopes;andiii) one region that specifically binds to a set of one or more Reck epitopes.

18. The antibody or antibody fragment according to any one of the previous claims 15 to 17, wherein the one or more binding domains of said antibody or antibody fragment are selected from one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains.

19. The antibody or antibody fragment according to any one of the previous claims 15 to 18, wherein said antibody or antibody fragment is a trispecific antibody with three binding domains, said binding domains including one Fzd binding domain, one Gprl24 binding domain, and one Reck binding domain.

20. The antibody or antibody fragment according to any one or the previous claims 15-19, wherein said antibody or antibody fragment does not comprise further binding domains over said one or more Fzd binding domains, one or more Gprl24 binding domains, and one or more Reck binding domains.

21. The antibody or antibody fragment according to any one of the previous claims 1-20, wherein said antibody or antibody fragment does not comprise an LRP binding domain such as an LRP5 or LRP6 binding domain, and / or is not able to bind to LRP such as an LRP5 or LRP6.

22. A nucleic acid encoding the antibody or antibody fragment according to any of the preceding claims 1 to 21 or a vector comprising said nucleic acid.

23. A pharmaceutical composition comprising a pharmaceutically acceptable excipient, diluent, or carrier, and the antibody or antibody fragment according to any of claims 1 to 21 or the nucleic acid or vector according to claim 22.

24. The antibody or antibody fragment according to any of claims 1-21, the nucleic acid according or the vector according to claim 22 for use as a medicament.

25. The antibody or antibody fragment according to any of claims 1-21, the nucleic acid according or the vector according to claim 22 for use in blood-brain barrier endothelial cell-directed gene therapy, or for use in the treatment of disorders linked to the blood-brain barrier integrity or for use in the treatment of the blood-brain barrier integrity.

26. The antibody or antibody fragment according to any of claims 1-21, the nucleic acid or the vector according to claim 22 for use in a treatment of a neurological disorder, said neurological disorder is preferably selected from the group of: retinopathy, ischemic stroke, hemorrhagic stroke, ischemia / reperfusion injury, brain aneurysms, arteriovenous malformations (AVMs), cavernous malformations, vasculitis, cerebral hemorrhage, subarachnoid hemorrhage, spinal vascular malformations, carotid artery stenosis, Moyamoya disease intracranial atherosclerosis, and combinations thereof, or multiple sclerosis, brain cancer, glioblastoma, human monogenic neurological disorders, neurodegenerative disorders, dementia, vascular dementia, HIV-l-associateddementia, Alzheimer's disease, Parkinson's disease, Huntington disease, amyotrophic lateral sclerosis, Charcot-Marie Tooth disease, dystonia, infectious brain diseases, traumatic brain injuries, migraine, neuroinflammation, COVID- 19, and chronic traumatic encephalopathy and combinations thereof.

27. The antibody or antibody fragment according to any of claims 1-21, the nucleic acid according or the vector according to claim 22 for use in blood-retina barrier endothelial cell-directed gene therapy, or for use in the treatment of disorders linked to the blood-retina barrier integrity or for use in the treatment of the blood-retina barrier integrity.

28. The antibody or antibody fragment according to any of claims 1-21, the nucleic acid or the vector according to claim 22 for use in a treatment of a ophthalmological disease or disorder, said ophthalmological disease or disorder is preferably selected from the group of: retinopathy, retinal vascular disorders such as Norrie disease, diabetic retinopathy, macular degeneration such as wet age-related macular degeneration and diabetic macular edema, familial exudative vitreoretinopathy, osteoporosis-pseudoglioma syndrome, retinal vein occlusion and retinopathy of prematurity.

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