Vectorized Anti-TDP-43 antibodies

Abstract

The present invention is in the field of AAV vectorized TDP-43 specific antibodies and uses thereof. The present invention further relates to means and methods to prevent, alleviate and / or treat a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, including but not limited to Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), Chronic Traumatic Encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

Description

[0001] VECTORIZED ANTI-TDP-43 ANTIBODIES

[0002] TECHNICAL FIELD

[0003] The invention relates to TDP-43 specific antibodies delivered by AAV (vectorized antibody) and uses thereof. The present invention provides means and methods to prevent, alleviate and / or treat a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, including but not limited to Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0004] BACKGROUND OF THE INVENTION

[0005] Age-associated brain disorders characterised by pathological aggregation of proteins in the central nervous system (CNS) (proteinopathies) and peripheral organs represent one of the leading causes of disability and mortality in the world. The best characterised protein that forms aggregates is amyloid beta in Alzheimer's disease and related disorders. Other disease- associated, aggregation-prone proteins leading to neurodegeneration include but are not limited to Tau, alpha-synuclein (aSyn, a-syn), huntingtin, fused in sarcoma (FUS), dipeptide repeat proteins (DPRs) produced by unconventional translation of the C9orf72 repeat expansion, superoxide dismutase 1 (SOD1), and TDP-43. Diseases involving TDP-43 aggregates are generally listed as TDP-43 proteinopathies including, but not limited to, ALS and FTD.

[0006] Transactive response (TAR) DNA binding protein 43 kDa (TDP-43) is a 414-amino acid protein encoded by the TARDBP gene on chromosome lp36.2 (ALS10). TARDBP is comprised of six exons (exon 1 is non-coding; exons 2-6 are protein-coding). TDP-43 belongs to the family of heterogeneous ribonucleoprotein (hnRNP) RNA binding proteins (Wang et al., Trends in Molecular Medicine Vol.14 No.11, 2008, 479-485; Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64). TDP-43 contains five functional domains (Figure 1 in Warraich et al., The International Journal of Biochemistry & Cell Biology 42 (2010) 1606-1609): two RNA recognition motifs (RRM1 and RRM2), which have two highly conserved hexameric ribonucleoprotein 2 (RNP2) and octameric ribonucleoprotein 1 (RNP1) regions, a nuclear export signal (NES) and a nuclear localization signal (NLS) enabling it to shuttle between the nucleus and the cytoplasm transporting bound mRNA, and a glycine- rich domain at the C-terminal, which mediates protein-protein interactions. TDP-43 is involved in multiple aspects of RNA processing, including transcription, splicing, transport, and stabilization (Buratti and Baralle, FEBS Journal 277 (2010) 2268-2281). It is a highly conserved, ubiquitously expressed protein with a tightly autoregulated expression level that shuttles continuously between the nucleus and cytoplasm. In normal physiological conditions, TDP-43 is predominantly localized in the nucleus. In 2006, TDP-43 was identified as the protein that accumulates in the vast majority of cases of frontotemporal lobar degeneration (FTLD) with tau-negative, ubiquitin-positive inclusions (then referred to as FTLD-TDP), and in most cases of amyotrophic lateral sclerosis (ALS) (Arai et al., Biochemical and Biophysical Research Communications 351 (2006) 602-611; Neumann et al., Science 314, (2006), 130- 133).

[0007] Thirty-eight negative-dominant mutations in TDP-43 have been identified in sporadic and familial ALS patients as well as in patients with inherited FTD mainly located in the glycine- rich domain (Figure 1 in Lagier-Tourenne and Cleveland, Cell 136, 2009, 1001-1004). TDP- 43 is inherently aggregation-prone, as shown by sedimentation assays, and this propensity is further increased by some of the ALS -associated TARDBP mutations (Ticozzi et al., CNS Neurol. Disord. Drug Targets. 2010, 9(3), 285-296.) connecting TDP-43 aggregation with clinical disease manifestation.

[0008] TDP-43 in neurodegeneration

[0009] TDP-43 aggregates have been identified in a growing list of neurodegenerative conditions (Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64), including but not limited to: Frontotemporal dementia (FTD, such as sporadic or familial with or without motor-neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutations, with TARDBP mutation, with valosin-containing protein (VCP) mutation, linked to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), Argyrophilic grain disease, Pick's disease, semantic variant Primary Progressive Aphasia (svPPA), behavioural variant FTD (bvFTD), nonfluent variant Primary Progressive Aphasia (nfvPPA) and the like), Amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD, including sporadic and familial forms of AD), Down syndrome, Familial British dementia, Polyglutamine diseases (Huntington’s disease and spinocerebellar ataxia type 3 (SC A3; also known as Machado Joseph Disease)), Hippocampal sclerosis dementia and Myopathies (sporadic inclusion body myositis, Inclusion body myopathy with a mutation in the valosin-containing protein ((VCP) and associated with Paget’s disease of the bone (PDB) and Frontotemporal dementia (FTD) abbreviated IBMPFD), Oculo-pharyngeal muscular dystrophy with rimmed vacuoles, Myofibrillar myopathies with mutations in the myotilin (MYOT) gene or mutations in the gene coding for desmin (DES)), Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s Disease (PD). The term LATE is intended to encompass several previously used designations related to TDP-43 proteinopathy that may be associated with cognitive impairment, including hippocampal sclerosis, hippocampal sclerosis of ageing, hippocampal sclerosis dementia, cerebral age-related TDP- 43 with sclerosis (CARTS), and TDP-43 pathologies in the elderly (for reviews see Kuslansky et al., 2004; Lippa and Dickson, 2004; Nelson et al., 2013, 2016b; Dutra et al., 2015).

[0010] Aggregated TDP-43 from patient brains shows a number of abnormal modifications, including hyperphosphorylation, ubiquitination, acetylation and C-terminal fragments through proteolytic cleavage (Arai et al., Biochemical and Biophysical Research Communications 351 (2006) 602-611; Neumann et al., Science 314, (2006), 130-133; Neumann et al., Acta Neuropathol. (2009) 117: 137-149; Hasegawa et al., (2008) Annals of Neurology Vol 64 No 1, 60-70; Cohen et al., Nat Commun. 6: 5845, 2015). Another characteristic feature of TDP- 43 pathology is redistribution and accumulation of TDP-43 from nucleus to cytoplasm. The hallmark lesions of FTLD-TDP are neuronal and glial cytoplasmic inclusions (NCI and GCI, respectively) and dystrophic neurites (DN) that are immunoreactive for TDP-43, as well as ubiquitin and p62, but negative for other neurodegenerative disease-related proteins. Differences in inclusion morphology and tissue distribution thereof are associated with specific mutations and / or clinical representations. Four types of TDP-43 pathology are described so far by histological classification (Mackenzie and Neumann, J. Neurochem. (2016) 138 (Suppl. 1), 54-70). FTLD-TDP type A cases are characterised by abundant short dystrophic neuritis (DN) and compact oval or crescentic NCI, predominantly in layer II of the neocortex (Fig. 2f in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Cases with this pathology usually present clinically with either behavioural -variant frontotemporal dementia (bvFTD) or nonfluent / agrammatic variants of Primary Progressive Aphasia (nfvPPA) and are associated with progranulin (GRN) mutations. Type B cases show moderate numbers of compact or granular NCI in both superficial and deep cortical layers with relatively few DN and Nil (neuronal intranuclear inclusions; Fig. 2g in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Most cases with co-appearance of FTD and ALS symptoms are found to have FTLD-TDP type B pathology. Type C cases have an abundance of long, tortuous neurites, predominantly in the superficial cortical laminae, with few or no NCI (Fig. 2j in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). This pathology is particularly found in cases presenting with semantic variant of Primary Progressive Aphasia (svPPA). FTLD-TDP type D displays with abundant lentiform neuronal intranuclear inclusions (Nil) and short DN in the neocortex with only rare NCI (Fig. 2k in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Type E is characterised by granulofilamentous neuronal inclusions (GFNIs) and very fine, dot-like neuropil aggregates affecting all neocortical layers in addition to curvilinear oligodendroglial inclusions in the white matter (Edward B. Lee et a / ., Acta Neuropathol. 2017 July; 134(1): 65-78.). This pattern of pathology is only found in cases with VCP in association with inclusion body myositis.

[0011] TDP-43 in ALS

[0012] Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterised by the premature loss of upper and lower motor neurons. The progression of ALS is marked by fatal paralysis and respiratory failure with a disease course from diagnosis to death of 1 to 5 years. In 97% of sporadic ALS, the neuropathology is characterised by abnormal cytoplasmic accumulations of TDP-43 in neurons and glia of the primary motor cortex, brainstem motor nuclei, spinal cord, and the associated white matter tracts (Prasad et al., 2019). ALS with dementia involves accumulation of TDP-43 in extramotor neocortex and hippocampus. The role of phosphorylation of TDP-43 in ALS patients has been explored with the help of antibodies that specifically bind to phosphorylated TDP-43 in nuclear and cytoplasmic inclusions with amino acids S379, S403, S404, S409, S410 as the major sites of phosphorylation of TDP-43 (Hasegawa et al., Ann Neurol 2008; 64: 60-70; Neumann et al., Acta Neuropathol (2009) 117: 137-149).

[0013] TDP-43 in FTD

[0014] Frontotemporal dementia (FTD) is a clinical term that covers a wide spectrum of disorders based on the degeneration of frontal and temporal lobes - a pathological feature termed frontotemporal lobar degeneration (FTLD). FTD is the second most abundant cause of early degenerative dementias in the age group below 65 years (Le Ber, Revue Neurologique 169 (2013) 811-819). FTD is presented by several syndromes including bvFTD which is characterised by changes in personality and behaviour; semantic dementia (SD) and progressive nonfluent aphasia (PNFA) characterised by changes in the language function; corticobasal syndrome (CBS), progressive supranuclear palsy syndrome and motor neuron disease (FTD-MND) characterised by movement dysfunction. Clinical diagnosis of these syndromes is complicated and final conclusion can only be achieved through postmortem histopathological analysis to detect aggregated protein and define affected brain regions. In terms of pathological, proteinaceous inclusions, about 45% of cases show pathological accumulation of misfolded Tau, 45% of cases have pathological TDP-43 and a smaller subgroup has aggregates of FUS and other proteins.

[0015] TDP-43 in AD and other diseases

[0016] TDP-43 pathology occurs in up to 57% of brains of patients with Alzheimer’s disease (Josephs KA et al., Acta Neuropathol. 2014; 127(6): 811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3): 441-450; McAleese et al., Brain Pathol. 2017 Jul; 27(4): 472-479). TDP-43 aggregation is associated with patient’s age and correlates with cognitive decline, memory loss and medial temporal atrophy in AD. It appears that in AD, TDP-43 represents a secondary or independent pathology that shares overlapping brain distribution with amyloid beta and tau pathologies in the medial temporal lobe. Pathologic TDP-43 follows a stereotypical pattern of progressive deposition that has been described by the so-called TDP-43 in AD (TAD) staging scheme: TDP-43 first deposits in the amygdala (stage I) followed by hippocampus, limbic, temporal, and finally the frontostriatum (stage V) (Josephs KA et al., Acta Neuropathol. 2014;127(6): 811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3): 441-450).

[0017] TDP-43 spreading

[0018] Although ALS and FTD onset and first symptoms vary significantly between patients, the common feature of disease progression is spreading of pathology from an initial focal area to most neurons. The continuous worsening of symptoms might be explained by the progressive spread of TDP-43 pathology. TDP-43 pathology in an ALS patient’s brain appears to be spreading in a four-stage process and it is believed that propagation occurs transynaptically via corticofugal axonal projections using anterograde axonal transport (Brettschneider et al., Ann Neurol. 2013 July; 74(1): 20-38.). Recent experimental evidence supports the hypothesis of protein propagation in neuronal tissue for amyloid-beta, Tau, alpha-synuclein and TDP-43 by a prion-like mechanism (Hasegawa et al., 2017), with starting points and the topographical spreading patterns being distinct for the four proteins (Brettschneider J et al., Nature Rev. Neuroscience, 2015, 109). The common, disease unifying mechanism is believed to be based on the cell-to-cell spreading of pathological protein aggregates. This mechanism consists of the release of aggregates from a diseased cell, uptake by a naive cell and seeding of the pathological protein conformation by a templated conformational change of endogenous proteins. Pathological TDP-43 able to induce aggregation of physiological (i.e. non-pathological TDP- 43) is defined as seeding-competent TDP-43. Indeed, TDP-43 has been found to misfold and aggregate into seeds that are propagating agents with the ability to trigger de novo misfolding. This “prion-like” paradigm is suspected to be one of the key elements in the disease progression.

[0019] TDP-43 cell-to cell spreading has been studied at a molecular level in few in vitro models, where insoluble TDP-43 preparations from patient brain are able to induce intracellular aggregate formation in receptor cells (Nonaka et al., Cell Reports 4 (2013), 124-134; Feiler et al., 2015; Porta et al., Nat. Comm., 2018). Further, it has been observed that intracellular TDP- 43 aggregates are released in association with exosome prior to spreading to the next cell (Nonaka et al., Cell Reports 4 (2013, 124-134)). Similarly, adenovirus-transduced TDP-43 expression led to cytoplasmic aggregates which were phosphorylated, ubiquitinated and more importantly acted as seeds initiating cell-to-cell spreading (Ishii et al., PLoS ONE 12(6): e0179375, 2017). The patient-derived pathological TDP-43 can lead to widespread deposition of endogenous TDP-43 following intracerebral inoculation into transgenic and wildtype mice (Porta et al., Nat. Comm., 2018).

[0020] Prevention and treatment of TDP-43 proteinopathies

[0021] TDP-43 aggregation and spreading of pathology are major hallmarks of ALS and FTD - fatal diseases for which currently no cure is available. To this end, anti-TDP-43 therapeutic antibodies have been developed in the last decade. WO2013 / 061163 describes TDP-43 specific binding molecules including polypeptides such as human antibodies as well as fragments, derivatives and variants thereof. WO2020 / 234473 describes TDP-43 specific binding molecules including polypeptides such as murine antibodies or antigen-binding fragments thereof. WO2023 / 156549 describes TDP-43 specific binding molecules including polypeptides such as humanized antibodies or antigen-binding fragments thereof. A major hurdle of using anti-TDP-43 antibodies in passive immunotherapy remains to achieve an effective antibody exposure in the central nervous system (CNS) in spite of the tightly regulated entry to the brain. Indeed, it has been reported that around 0.1% of an injected antibody reaches the brain following peripheral administration (St-Amour et al., 2013; Wang et al., 2018; Pardridge et al., 2019). Achieving a sufficient concentration of antibody in the brain to elicit a therapeutic response remains a challenge, even with high injected dose. Such drawback has been described as one of the possible causes of failure of passive immunotherapies during clinical trials (Thom et al., 2018). In recent years, vectorized delivery of antibodies has emerged as a potential approach for antibody delivery to the CNS. WO2017 / 189963 describes vectorized delivery of anti-Tau antibody and AAV vectors expression cassettes. Tamaki et al., 2018 reports vectorized delivery of an anti-TDP-43 scFv. Patent application WO2022 / 129609 describes vectorized delivery of anti-TDP-43 antibody and AAV vectors expression cassettes.

[0022] SUMMARY OF THE INVENTION

[0023] At present there are no approved therapies on the market to treat and / or prevent TDP-43 associated diseases. There is therefore a pressing need to identify new therapies that can treat and / or prevent these diseases. Specifically related to the vectorized approach, the further development and optimization of the vectorized delivery of anti-TDP-43 antibody are desirable in order to address the unmet need for methods of treatment and prevention of diseases associated with TDP-43. Accordingly, in a first aspect, the invention provides an adeno- associated virus (AAV) vector comprising a nucleic acid encoding an anti-TDP-43 antibody. The AAV vector comprises an expression cassette comprising a nucleic acid encoding from 5’ to 3’ : a promoter (that may be ubiquitous or CNS specific), a light chain of an anti-TDP-43 antibody, an internal ribosome entry site (IRES), a heavy chain of an anti-TDP-43 antibody and a regulatory element. More specifically, the invention provides an AAV vector comprising an expression cassette comprising a nucleic acid encoding an anti-TDP-43 antibody. The AAV vector comprises an expression cassette comprising a nucleic acid encoding from 5’ to 3’: a promoter, a light chain of an antibody, an internal ribosome entry site (IRES), a heavy chain of an antibody and a regulatory element; wherein the light chain of the antibody comprises a secretion peptide and / or the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17. The invention also provides a pharmaceutical composition comprising an AAV vector of the invention and a pharmaceutically acceptable carrier and / or excipient and / or diluent.

[0024] The invention also provides an AAV vector of the invention or pharmaceutical composition of the invention for use in a method of reducing the level of phosphorylated TDP-43 in a subject.

[0025] The invention also provides an AAV vector of the invention or pharmaceutical composition of the invention for use in prevention, alleviation or treatment of a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy.

[0026] The invention also provides a method of retaining or increasing cognitive memory capacity or slowing memory loss in a subject with a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, comprising administering an AAV vector or pharmaceutical composition of the invention to the subject.

[0027] The invention also provides a method of reducing the level of pathological TDP-43, in particular the level of aggregated TDP-43 and / or phosphorylated TDP-43, in a subject, comprising administering an AAV vector or pharmaceutical composition of the invention to the subject.

[0028] The invention also provides methods of inducing production and / or sustained expression of an anti-TDP-43 antibody in a subject, comprising administering an AAV vector (as described herein) to the subject.

[0029] The invention also provides a method for producing an AAV vector, comprising: (i) providing a host cell comprising the viral genome of the AAV vector described herein (ii) incubating the host cell under conditions suitable to enclose the viral genome in an AAV capsid, thereby making the AAV vector.

[0030] DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

[0031] Definitions

[0032] Antibody The terms "anti-TDP-43 antibody" or "an antibody that binds to TDP-43", as used herein refer to an antibody that is capable of binding TDP-43 with sufficient affinity such that the antibody is considered for further assessment as a potential therapeutic agent in targeting TDP-43. In general, the term "antibody" is used herein in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multi-specific antibodies (e.g., bispecific or biparatopic antibodies), fully-human antibodies and antibody fragments so long as they exhibit the desired antigen-binding activity. Antibodies within the present invention may also be chimeric antibodies, recombinant antibodies, antigenbinding fragments of recombinant antibodies, human antibodies, or humanized antibodies. Antibodies within the present invention may also include “vectorized antibodies” as defined elsewhere herein.

[0033] An "antigen-binding fragment" or “functional fragment thereof’ refers to a molecule other than an intact, or full-length, antibody that comprises a portion of an intact, or full-length, antibody and that binds (fully or partially) the antigen to which the intact, or full-length, antibody binds. Examples of antigen-binding fragments include but are not limited to Fv, Fab, Fab', Fab' -SH, F(ab')2; diabodies; linear antibodies; single domain antibody, single-chain antibody molecules (e.g. scFv); and multi-specific antibodies formed from antibody fragments. Antigen-binding fragments may also be referred to as “functional fragments” as they retain the binding function of the original antibody from which they are derived. For the avoidance of doubt, the term "anti-TDP-43 antibody" as defined above encompasses antigen-binding fragments and functional fragments thereof.

[0034] An "antibody that binds to an epitope" within a defined region of a protein is an antibody that requires the presence of one or more of the amino acids within that region for binding to the protein.

[0035] The term “binding to” as used in the context of the present invention defines a binding (interaction) of at least two “antigen-interaction-sites” with each other. The term “antigen- interaction-site” defines, in accordance with the present invention, a motif of a polypeptide, i.e., a part of the antibody or antigen-binding fragment of the present invention, which shows the capacity of specific interaction with a specific antigen or a specific group of antigens of TDP-43. Said binding / interaction is also understood to define a “specific recognition”. The term “specifically recognizing” means in accordance with this invention that the antibody is capable of specifically interacting with and / or binding to at least two amino acids of TDP-43 as defined herein, in particular interacting with / binding to at least two amino acids within amino acids residues 397-411 of human TDP-43 (SEQ ID NO: 1), even more particularly interacting with binding to at least two amino acids within amino acids residues 400-405, 400- 406 or 400-412 of human TDP-43 (SEQ ID NO: 1).

[0036] The term “specific interaction” as used in accordance with the present invention means that the antibody or antigen-binding fragment thereof of the invention does not or does not essentially cross-react with (poly)peptides of similar structures. Accordingly, the antibody or antigenbinding fragment thereof of the invention specifically binds to / interacts with structures of TDP- 43 formed by particular amino acid sequences within amino acids residues 397-411 of human TDP-43 (SEQ ID NO: 1), more particularly binds to / interacts with structures of TDP-43 formed by particular amino acid sequences within amino acids residues 400-405, 400-406 or 400-412 of human TDP-43 (SEQ ID NO: 1).

[0037] Cross-reactivity of antigen-binding molecules, in particular a panel of antibodies or antigenbinding fragments thereof under investigation may be tested, for example, by assessing binding of said panel of antibodies or antigen-binding fragments thereof under conventional conditions (see, e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1988) and Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1999)) to the (poly)peptide of interest as well as to a number of more or less (structurally and / or functionally) closely related (poly)peptides. Only those constructs (i.e. antibodies, antigen-binding fragments thereof and the like) that bind to the certain structure of TDP-43 as defined herein, e.g., a specific epitope or (poly)peptide / protein of TDP-43 as defined herein but do not or do not essentially bind to any of the other epitope or (poly )pepti des of the same TDP-43, are considered specific for the epitope or (poly)peptide / protein of interest and selected for further studies in accordance with the method provided herein. These methods may comprise, inter alia, binding studies, blocking and competition studies with structurally and / or functionally closely related molecules. These binding studies also comprise FACS analysis, surface plasmon resonance (SPR, e.g. with BIACORE™), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy or by radiolabeled ligand binding assays. Accordingly, specificity can be determined experimentally by methods known in the art and methods as described herein. Such methods comprise, but are not limited to Western Blots, ELISA-, RIA-, ECL-, IRMA-tests and peptide scans.

[0038] The term “monoclonal antibody” as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, z.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Monoclonal antibodies are advantageous in that they may be synthesized by a hybridoma culture, essentially uncontaminated by other immunoglobulins. The modified "monoclonal" indicates the character of the antibody as being amongst a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. As mentioned above, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method described by Kohler, Nature 256 (1975), 495.

[0039] The term “polyclonal antibody” as used herein, refers to an antibody which was produced among or in the presence of one or more other, non-identical antibodies. In general, polyclonal antibodies are produced from a B-lymphocyte in the presence of several other B-lymphocytes which produced non-identical antibodies. Usually, polyclonal antibodies are obtained directly from an immunized animal.

[0040] The term “fully-human antibody” as used herein refers to an antibody which comprises human immunoglobulin protein sequences only. A fully human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “murine antibody” refers to an antibody which comprises mouse / murine immunoglobulin protein sequences only. Alternatively, a “fully- human antibody” may contain rat carbohydrate chains if produced in a rat, in a rat cell, in a hybridoma derived from a rat cell. Similarly, the term “rat antibody” refers to an antibody that comprises rat immunoglobulin sequences only. Fully-human antibodies may also be produced, for example, by phage display which is a widely used screening technology which enables production and screening of fully human antibodies. Also, phage antibodies can be used in context of this invention. Phage display methods are described, for example, in US 5,403,484, US 5,969,108 and US 5,885,793. Another technology which enables development of fully- human antibodies involves a modification of mouse hybridoma technology. Mice are made transgenic to contain the human immunoglobulin locus in exchange for their own mouse genes (see, for example, US 5,877,397).

[0041] The term “chimeric antibodies”, refers to an antibody which comprises a variable region of the present invention fused or chimerized with an antibody region (e.g., constant region) from another, human or non-human species (e.g., mouse, horse, rabbit, dog, cow, chicken).

[0042] The term antibody also relates to recombinant human antibodies, heterologous antibodies and heterohybrid antibodies. The term "recombinant (human) antibody" includes all human sequence antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes; antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant, combinatorial human antibody library, or antibodies prepared, expressed, created or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions (if present) derived from human germline immunoglobulin sequences. Such antibodies can, however, be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.

[0043] A "heterologous antibody" is defined in relation to the transgenic non-human organism producing such an antibody. This term refers to an antibody having an amino acid sequence or an encoding nucleic acid sequence corresponding to that found in an organism not consisting of the transgenic non-human animal, and generally from a species other than that of the transgenic non-human animal.

[0044] The term "heterohybrid antibody" refers to an antibody having light and heavy chains of different organismal origins. For example, an antibody having a human heavy chain associated with a murine light chain is a heterohybrid antibody. Examples of heterohybrid antibodies include chimeric and humanized antibodies. In one aspect, the invention relates to humanized antibodies. "Humanized" forms of non-human (e.g. murine or rabbit) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding sub-sequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Often, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibody may comprise residues, which are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see: Jones et al., Nature 321 (1986), 522-525; Reichmann Nature 332 (1998), 323-327 and Presta Curr Op Struct Biol 2 (1992), 593-596.

[0045] Accordingly, in the context of the present invention, the term “antibody” relates to full immunoglobulin molecules as well as to parts of such immunoglobulin molecules (i.e., “antigen-binding fragment thereof’). Furthermore, the term relates, as discussed above, to modified and / or altered antibody molecules. The term also relates to intact antibodies as well as to antibody fragments thereof, like, separated light and heavy chains, Fab, Fv, Fab’, F(ab’)2. The term antibody also comprises but is not limited to fully-human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies and antibody constructs, like antibody-fusion proteins.

[0046] The "Fv region" comprises the variable regions from both the heavy and light chains but lacks the constant regions. The fragment antigen-binding (“Fab fragment”) is a region of an antibody that binds to antigens. It is composed of one constant and one variable domain of each of the heavy and the light chain. A Fab fragment contains an interchain disulfide bond that link the constant domain of the light chain and the constant domain of the heavy chain.

[0047] A "Fab1fragment" is composed of one constant and one variable domain of each of the heavy and the light chain. A Fab fragment contains an interchain disulfide bond that link the light chain and the heavy chain. It also contains the region between the CHI and CH2 domains, such that an interchain disulfide bond can be formed between the two heavy chains of two Fab' fragments to form a F(ab')2 molecule.

[0048] A "F(ab')2 fragment" contains two light chains and two heavy chains containing a portion of the constant region between the CHI and CH2 domains, such that an interchain disulfide bond is formed between the two heavy chains. A F(ab')2 fragment is composed of two Fab' fragments that are held together by a disulfide bond between the two heavy chains.

[0049] An "Fc" region contains two heavy chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains.

[0050] Humanized antibodies, humanized antibody constructs, humanized antibody fragments, humanized antibody derivatives (all being Ig-derived) to be employed in accordance with the invention or their corresponding immunoglobulin chain(s) can be further modified using conventional techniques known in the art, for example, by using amino acid deletion(s), insertion(s), substitution(s), addition(s), and / or recombination(s) and / or any other modification(s) known in the art either alone or in combination. Methods for introducing such modifications in the DNA sequence underlying the amino acid sequence of an immunoglobulin chain are well known to the person skilled in the art; see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2ndedition (1989) and 3rdedition (2001). The term “Ig-derived domain” particularly relates to (poly)peptide constructs comprising at least one CDR. Fragments or derivatives of the recited Ig-derived domains define (poly)peptides which are parts of the above antibody molecules and / or which are modified by chemical / biochemical or molecular biological methods. Corresponding methods are known in the art and described inter alia in laboratory manuals (see Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2nd edition (1989) and 3rd edition (2001); Gerhardt et al., Methods for General and Molecular Bacteriology ASM Press (1994); Lefkovits, Immunology Methods Manual: The Comprehensive Sourcebook of Techniques; Academic Press (1997); Golemis, Protein-Protein Interactions: A Molecular Cloning Manual Cold Spring Harbor Laboratory Press (2002)).

[0051] The term “CDR” as employed herein relates to “complementary determining region”, which is well known in the art. The CDRs are parts of immunoglobulins that determine the specificity of said molecules and make contact with a specific ligand. The CDRs are the most variable part of the molecule and contribute to the diversity of these molecules. There are three CDR regions CDR1, CDR2 and CDR3 in each V domain. CDR-H depicts a CDR region of a variable heavy chain and CDR-L relates to a CDR region of a variable light chain. VH means the variable heavy chain and VL means the variable light chain. The CDR regions of an Ig-derived region may be determined as described in Kabat “Sequences of Proteins of Immunological Interest”, 5th edit. NTH Publication no. 91-3242 U.S. Department of Health and Human Services (1991). CDR sequences provided herein are defined according to Kabat. However, it will be understood by the skilled person that the invention is intended to encompass binding molecules in which the CDR sequences are defined according to any useful identification / numbering scheme. For example, Chothia (Canonical structures for the hypervariable regions of immunoglobulins. Chothia C, Lesk AM. J Mol Biol. 1987 Aug 20; 196(4):901-17), IMGT (IMGT, the international ImMunoGeneTics database. Giudicelli V, Chaume D, Bodmer J, Muller W, Busin C, Marsh S, Bontrop R, Marc L, Malik A, Lefranc MP. Nucleic Acids Res. 1997 Jan 1; 25(l):206-l 1 and Unique database numbering system for immunogenetic analysis. Lefranc MP. Immunol Today. 1997 Nov; 18(11):509), MacCallum (MacCallum RM, Martin AC, Thornton JM, J Mol Biol. 1996 Oct 11; 262(5):732-45) and Martin (Abhinandan KR, Martin ACR. Analysis and improvements to Kabat and structurally correct numbering of antibody variable domains. Mol Immunol. (2008) 45:3832-9. 10.1016 / j.molimm.2008.05.022) numbering schemes may be adopted in order to define the CDRs.

[0052] “Humanization approaches” are well known in the art and in particular described for antibody molecules, e.g. Ig-derived molecules. The term “humanized” refers to humanized forms of nonhuman (e.g., murine) antibodies or fragments thereof (such as Fv, Fab, Fab’, F(ab’), scFvs, or other antigen-binding partial sequences of antibodies) which contain some portion of the sequence derived from non-human antibody. Humanized antibodies include human immunoglobulins in which residues from a complementary determining region (CDR) of the human immunoglobulin are replaced by residues from a CDR of a non-human species such as mouse, rat or rabbit having the desired binding specificity, affinity and capacity. In general, the humanized antibody will comprise substantially all of at least one, and generally two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non- human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin ; see, inter alia, Jones et al., Nature 321 (1986), 522-525, Presta, Curr. Op. Struct. Biol. 2 (1992), 593-596. Methods for humanizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acids introduced into it from a source which is non-human still retain the original binding activity of the antibody. Methods for humanization of antibodies / antibody molecules are further detailed in Jones et al., Nature 321 (1986), 522-525; Reichmann et al., Nature 332 (1988), 323-327; and Verhoeyen et al., Science 239 (1988), 1534-1536. Specific examples of humanized antibodies, e.g. antibodies directed against EpCAM, are known in the art (see e.g. LoBuglio, Proceedings of the American Society of Clinical Oncology Abstract (1997), 1562 and Khor, Proceedings of the American Society of Clinical Oncology Abstract (1997), 847).

[0053] In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgGl, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.

[0054] In certain embodiments, the antibody provided herein binds to pathological TDP-43 and forms an immune complex that may be cleared by antibody-dependent cellular phagocytosis (ADCP), which as a result potentiates TDP-43 clearance. ADCP is mediated by the interaction of antibody Fc fragment with Fc receptors, such as Fc gamma receptors, expressed at the surface of innate immune cells, such as microglia or dendritic cells. Fc mediated functions may be modulated to achieve the desired effect by modifying the Fc portion of antibodies.

[0055] In certain embodiments, the invention contemplates an antibody variant that possesses some but not all effector functions, which make it a desirable candidate for applications in which the half-life of the antibody in vivo is important yet certain effector functions (such as complement activation and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be conducted to confirm the reduction / depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) binding assays can be conducted to ensure that the antibody lacks FcyR binding (hence likely lacking ADCC activity), but retains FcRn binding ability. The primary cells for mediating ADCC, NK cells, express FcyRIII only, whereas monocytes and microglia express FcyRI, FcyRII and FcyRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Patent No. 5,500,362 (see, e.g. Hellstrom, I. et al., Proc. Nat’l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat’l Acad. Sci. USA 82: 1499- 1502 (1985); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166: 1351-1361 (1987)).

[0056] Alternatively, non-radioactive assays methods may be employed (see, for example, ACTI™ non radioactive cytotoxicity assay for flow cytometry (Cell Technology, Inc. Mountain View, CA; and CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.

[0057] Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al., Proc. Nat'lAcad. sci. USA 95:652-656 (1998).

[0058] Clq binding assays may also be carried out to confirm that the antibody is unable to bind Clq and hence lacks CDC activity. See, e.g., Clq and C3c binding ELISA in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano- Santoro et al., J. Immunol. Methods 202: 163 (1996); Cragg, M.S. et al., Blood 101 : 1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life determinations can also be performed using methods known in the art (see, e.g., Petkova, S.B. et al., Int'L Immunol. 18(12): 1759- 1769 (2006)). Antibodies with reduced effector function include those with substitution of one or more of Fc region residues 234, 235, 238, 265, 269, 270, 297, 327 and 329 (U.S. Patent No. 6,737,056). Certain antibody variants with improved or diminished binding to FcRs are described. (See, e.g., U.S. Patent No. 6,737,056; WO 2004 / 056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" Fc mutant with substitution of residues 265 and 297 to alanine (US Patent No. 7,332,581) or the so-called “DANG” Fc mutant with substitution of residues 265 to alanine and 297 to glycine. Alternatively, antibodies with reduced effector function include those with substitution of one or more of Fc region residues 234, 235 and 329, so-called “PG-LALA” Fc mutant with substitution of residues 234 and 235 to alanine and 329 to glycine (Lo, M. et al., Journal of Biochemistry, 292, 3900-3908). Other known mutations at position 234, 235 and 321, the so- called TM mutant containing mutations L234F / L235E / P331 S in the CH2 domain, can be used (Oganesyan et al. Acta Cryst. D64, 700-704. (2008)). Antibodies from the human IgG4 isotype include mutations S228P / L235E to stabilize the hinge and to reduce FgR binding (Schlothauer et al, PEDS, 29 (10):457-466). Numbering of the constant domain is according to the EU numbering system. In one embodiment, the anti-TDP-43 antibody comprises an S228P mutation.

[0059] Other Fc variants include those with substitutions at one or more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (US Patent No. 7,371,826). See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821.

[0060] In certain embodiments, the Fc region is mutated to increase its affinity to FcRn at pH 6.0 and consequently extend the antibody half-life. Antibodies with enhanced affinity to FcRn include those with substitution of one or more of Fc region residues 252, 253, 254, 256, 428, 434, including the so called YTE mutation with substitution M252Y / S254T / T256E (DalF Acqua et al, J Immunol. 169:5171-5180 (2002)) or LS mutation M428L / N434S (Zalevsky et al, Nat Biotechnol. 28(2): 157-159 (2010)).

[0061] The term "clearance” (also referred as “clearance value” or “CL” or “systemic clearance”) relates to the efficiency of elimination of a substance from the body. Clearance of a substance (in this case a binding molecule of the invention) is the sum of the urinary and extrarenal clearance; for substances that are eliminated by renal and extrarenal routes, plasma clearance exceeds urinary clearance. The PK properties of mAbs are a function of their large size (150 kDa), relative polarity, Fc-receptor binding and specific binding to target antigens. The primary elimination route for mAbs is cellular uptake followed by proteolytic degradation. Low clearance of mAbs from the systemic circulation enables them to be administered less frequently than peptides or small molecules, which is often more convenient for patients (Betts et al., MABs. 2018).

[0062] Vectorized antibody

[0063] The term “vector” refers to any particle used to transport (by transduction or transfection) a polynucleotide(s) into a host cell. This definition includes both non-viral and viral vectors.

[0064] The term “viral vector” includes vectors derived from wild-type viruses and engineered (e.g. modified) viruses. Examples of viral vectors include, but are not limited to, adeno associated virus (AAV), adenovirus, retrovirus, rhinovirus, lentivirus, hepatovirus, herpes simplex virus (HSV) and any virus-like particle.

[0065] The invention is directed to using AAV vectors to deliver TDP-43 specific antibodies. As used herein, an “AAV vector” refers to an adeno-associated virus suitably used to transport a polynucleotide(s) into a host cell. The AAV can be of any suitable serotypes, examples of which include, but are not limited to, AAV serotype 1 (AAV1), AAV serotype 2 (AAV2), AAV serotype 3 (AAV3), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5), AAV serotype 6 (AAV6), AAV serotype 7 (AAV7), AAV serotype 8 (AAV8), AAV serotype 9 (AAV9), AAV serotype 10 (AAV 10), AAV serotype 11 (AAV 11), or AAV serotype 12 (AAV 12), or any other wild-type serotypes or engineered AAVs.

[0066] According to the invention, a “vectorized antibody” refers to an antibody delivered as a corresponding nucleic acid molecule encoding said antibody. Such nucleic acid molecule is comprised in an AAV vector, for targeted delivery to host cells. In other words, a “vectorized antibody” is used herein to refer to the resulting antibody that is the expression product of the AAV vector comprising a nucleic acid molecule (in the form of an expression cassette) that encodes an anti-TDP-43 antibody. As used herein, an “expression cassette” refers to a nucleic acid construct comprising various operably linked polynucleotide elements, including a gene destined for expression. In accordance with the present invention, the expression cassette comprises at least two genes for expression: one encoding the light chain and one encoding the heavy chain of an anti-TDP-43 antibody. The expression cassette may comprise sequences providing or encoding one or more of a promoter operably linked to the nucleic acid encoding an antibody, a ribosomal binding site, a start codon, a stop codon, and a transcription termination sequence. Suitably the expression cassette may further comprise a nucleic acid encoding a post-transcriptional regulatory element. Suitably the expression cassette may additionally comprise a nucleic acid encoding a poly A (polyadenylation) element.

[0067] The term “operably linked” as used herein refers to the arrangement of various nucleic acid elements relative to each such that the elements are functionally connected and are able to interact with each other in the manner intended. Such elements may include, without limitation, a promoter, an enhancer and / or a regulatory element, a polyadenylation element, one or more introns and / or exons, and a coding sequence of a gene or genes of interest to be expressed. The nucleic acid elements, when properly oriented or operably linked, act together to modulate the activity of one another, and ultimately may affect the level of expression of a product (e.g. an antibody). By “modulate” is meant increasing, decreasing, or maintaining the level of activity of a particular element. As understood by a person of average skill in the art, “operably linked” implies functional activity, and is not necessarily related to a natural positional link.

[0068] As used herein, the term “regulatory element” refers to a nucleic acid sequence capable of modulating (e.g. increasing or decreasing) the expression of an operably linked sequence (e.g. a gene). Non-limiting examples of regulatory elements include promoter, enhancer, repressor, silencer, intron, UTR, miR binding sites, post-translational response element, post- transcriptional regulatory element (e.g. WPRE) and poly A element.

[0069] As used herein, the term "promoter" refers to a region of DNA that generally is located upstream of a polynucleotide sequence (e.g. the polynucleotide sequence encoding the light chain and heavy chain of an anti-TDP-43 antibody) to be transcribed that is needed for transcription to occur, e.g. which initiates transcription. Non-limiting examples of promoters include cytomegalovirus (CMV) promoter, EFl A (Human Eukaryotic translation elongation factor 1 alpha 1), CAG (CMV early enhancer fused to modified chicken P-actin promoter), CBA (CMV early enhancer fused to chicken P-actin promoter and SV40 intron), CBh (CMV early enhancer fused to modified chicken P-actin promoter), SV40 (Simian virus 40 enhancer / early promoter), GfaABCID (modified GFAP promoter), CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), ATP1A2 (Na, K ATPase a2), CLDN5 (Claudin 5), ADRB2 (Adrenoceptor beta 2), TNFRSF6B (TNF receptor superfamily member 6b), PDYN (prodynorphin), GH1 (Human growth hormone), OPALIN (Opalin), SYN1 (Synapsin 1), CAMK2A (Calcium / Calmodulin Dependent Protein Kinase II alpha), NEFH (neurofilament heavy polypeptide), NEUROD6 (neuronal differentiation factor 6), OLIG2 (oligodendrocyte transcription factor 2) or CMVe- OLIG2 (CMV enhancer fused to OLIG2 promoter).

[0070] The term promoter includes synthetic promoters. The term “synthetic promoter” as used herein relates to a promoter that does not occur in nature. For example, functional variants of naturally occurring promoters can be used in accordance with the invention. A “functional variant” of a promoter in the context of the present invention is a variant of a reference promoter that retains the ability to function in the same way as the reference promoter. In additional embodiments, truncated forms of naturally occurring promoters are used. In some embodiments, the promoter is operably linked to an enhancer such as the CMV early enhancer, or fused to an intron sequence(s). Truncated or modified naturally occurring promoters can be used to facilitate insertion of relatively large antibody encoding sequences into a vector, in particular a viral vector.

[0071] The expression cassette of the invention also comprises a sequence encoding an “internal ribosome entry site” (IRES). As used herein, the term “internal ribosome entry site” refers to an RNA element that acts to promote recruitment of ribosomes. The IRES element is therefore able to promote protein translation in a 5’ cap independent manner. An IRES element is advantageous in expression cassettes containing more than one gene for expression (in this case, separate expression of light and heavy chains).

[0072] The AAV vector as described herein may specifically target cells of the central nervous system (CNS) or the blood-brain barrier (BBB). However, in other instances the AAV vector does not specifically target the CNS or the BBB. For example, many wild-type viral vectors target any tissue or cell-type and thus include the CNS or the BBB. In such cases, CNS-specific or BBB- specific promoters can be used to drive the expression of the polynucleotide encoding an antibody or antibody fragment in the cells of the CNS or the BBB in a preferential or predominant manner as compared to other tissues.

[0073] The AAV vector may comprise an expression cassette comprising a nucleic acid sequence encoding for a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) at the 3 ‘end. The WPRE sequence is routinely used to increase expression of genes delivered by viral vectors. Without wishing to be bound by theory, inclusion of the sequence in expression cassettes can increase mRNA stability and thus protein yield.

[0074] The expression cassette of the invention also comprises sequences encoding at least one secretion peptide at the 5’ terminus of the antibody light chain and / or heavy chain . A “secretion peptide” as used herein (also referred to interchangeably as signal peptide / sequence, targeting signal, localization signal / sequence, transit peptide, leader sequence / peptide) refers to a short peptide, generally under 30 amino acids and usually located at the N-terminus of newly synthesized proteins, that are destined toward the secretory pathway. The at least one secretion peptide may assist with delivery and secretion of the antibody into the CNS. In instances in which the expression cassette encodes multiple genes, all genes may further comprise a sequence encoding a secretion peptide. The light chain and / or heavy can comprise a secretion peptide. This may be alternatively specified that the light chain and / or heavy chain can be preceded by a secretion peptide. In other words, the secretion peptide can be considered located at the 5’ terminus preceding the light chain and / or heavy chain. The secretion peptide typically comprises a charged N-terminal region, a hydrophobic core region and a C-terminal region recognised by a signal peptidase. Non limiting examples of secretion peptides are described in O’Neill et al., 2023.

[0075] The AAV vectors as described herein can be administered to the subject by any conventional route, including injection or by gradual infusion over time. The administration may be via parenteral administration. The administration may, for example, be by infusion or by intravenous, intraperitoneal, intranasal, intravitreous, subcutaneous, intramuscular, intrathecal, intraci sternal, intraparenchymal, intrastriatal or intracerebroventricular route. As further examples, suitable forms for parenteral injection (including, subcutaneous, intramuscular, intravascular or infusion) include a sterile solution, suspension or emulsion. The identification of suitable dosages of the compositions of the invention is well within the routine capabilities of a person of average skill in the art. For example, the suitable dosage for a given subject will be determined by taking into consideration various factors known to modify the action of the vector for the uses according to the invention. For example, severity and type of disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, body weight, sex, diet, time and route of administration, other medications and other relevant clinical factors. The dosages and schedules may be varied according to the particular condition, disorder or symptom the overall condition of the subject. It may also be the case that there is no single accepted dose for the treatment of a given disease, but that a range of doses is considered suitable. Effective dosages may be determined by either in vitro or in vivo methods.

[0076] Pharmaceutically acceptable carriers, diluents, adjuvants and excipients are well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, 15th or 18th Ed. (Alfonso R. Gennaro, ed.; Mack Publishing Company, Easton, PA, 1990); Remington: the Science and Practice of Pharmacy 19th Ed. (Lippincott, Williams & Wilkins, 1995); Handbook of Pharmaceutical Excipients, 3rd Ed. (Arthur H. Kibbe, ed.; Amer. Pharmaceutical Assoc, 1999); Pharmaceutical Codex: Principles and Practice of Pharmaceutics 12th Ed. (Walter Lund ed.; Pharmaceutical Press, London, 1994); The United States Pharmacopeia: The National Formulary (United States Pharmacopeial Convention); Fiedler’s “Lexikon der Hilfstoffe” 5th Ed., Edition Cantor Verlag Aulendorf 2002; “The Handbook of Pharmaceutical Excipients”, 4th Ed., American Pharmaceuticals Association, 2003; and Goodman and Gilman's: the Pharmacological Basis of Therapeutics (Louis S. Goodman and Lee E. Limbird, eds.; McGraw Hill, 1992), the disclosures of which are hereby incorporated by reference.

[0077] The carriers, diluents, adjuvants and pharmaceutical excipients can be selected with regard to the intended route of administration and standard pharmaceutical practice. These compounds must be acceptable in the sense of being not deleterious to the recipient thereof. See Remington's Pharmaceutical Sciences, 15th or 18th Ed. (Alfonso R. Gennaro, ed.; Mack Publishing Company, Easton, PA, 1990); Remington: the Science and Practice of Pharmacy 19th Ed. (Lippincott, Williams & Wilkins, 1995); Handbook of Pharmaceutical Excipients, 3rd Ed. (Arthur H. Kibbe, ed.; Amer. Pharmaceutical Assoc, 1999); Pharmaceutical Codex: Principles and Practice of Pharmaceutics 12th Ed. (Walter Lund ed.; Pharmaceutical Press, London, 1994); The United States Pharmacopeia: The National Formulary (United States Pharmacopeial Convention); Fiedler’s “Lexikon der Hilfstoffe” 5th Ed., Edition Cantor Verlag Aulendorf 2002; “The Handbook of Pharmaceutical Excipients”, 4th Ed., American Pharmaceuticals Association, 2003; and Goodman and Gilman's: the Pharmacological Basis of Therapeutics (Louis S. Goodman and Lee E. Limbird, eds.; McGraw Hill, 1992), the disclosures of which are hereby incorporated by reference. Carriers, diluents, adjuvants and pharmaceutical excipients for AAV formulations are known in the art such as described and referenced in Chan et al., 2022.

[0078] The "effective amount" of the compound which is to be administered to a subject is the dosage which according to sound medical judgement is suitable for treating, preventing or alleviating the disease, disorder or abnormality. The specific dose level and frequency of dosage can depend, e.g., upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, mode and time of administration, the rate of excretion, and drug combination. Patient-specific factors such as the age, body weight, general health, sex, diet, as well as the severity of the particular condition can also influence the amount which is to be administered.

[0079] Invention embodiments

[0080] In one aspect of the invention, there is provided an AAV vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a regulatory element, wherein the light chain of the antibody comprises a secretion peptide and / or the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0081] The AAV vector may comprise an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’: (i) a promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a regulatory element, wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0082] In some embodiments, the first and second secretion peptide are the same.

[0083] In a further aspect the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a promoter, (ii) a first secretion peptide, (iii) the light chain of an antibody, (iv) an internal ribosome entry site (IRES), (v) a second secretion peptide, (vi) the heavy chain of an antibody, and (vii) a regulatory element, wherein the antibody is an anti- TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0084] In some embodiments, the first and second secretion peptide are the same.

[0085] In some embodiments, the AAV vector comprises an anti-TDP-43 antibody comprising a VH- CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17. In an embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 or a VH having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 14 or a VL having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14. In an embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 and a VL comprising the amino acid sequence of SEQ ID NO: 14. The anti-TDP-43 antibody may be a murine or chimeric antibody.

[0086] In some embodiments, the AAV vector comprises an anti-TDP-43 antibody comprising a VH- CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27. In an embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 or a VH having at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a VL comprising the amino acid sequence of SEQ ID NO: 24 or a VL having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24. In an embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 and a VL comprising the amino acid sequence of SEQ ID NO: 24. The anti-TDP-43 antibody may be a humanized antibody.

[0087] In some embodiments, the AAV vector is single stranded. In some embodiments, the AAV vector is self-complementary. In some embodiments, the AAV vector is codon-optimized.

[0088] As noted above, the AAV vector of the invention comprises an expression cassette, wherein the expression cassette comprises a promoter. In some embodiments, the promoter is a ubiquitous promoter. A “ubiquitous” promoter is a promoter that drives expression of the target genes (i.e. the light chain and heavy chain of an anti-TDP-43 antibody) in a range of different tissues and cell types. Examples of ubiquitous promoters are the CBh promoter (CMV early enhancer fused to modified chicken P-actin promoter), EFl A promoter (Human Eukaryotic translation elongation factor 1 alpha 1), CAG promoter (CMV early enhancer fused to modified chicken P-actin promoter), CBA promoter (CMV early enhancer fused to chicken P-actin promoter and SV40 intron) or SV40 promoter (Simian virus 40 enhancer / early promoter). In other embodiments, the promoter is a CNS-specific promoter. A “CNS-specific” promoter is a promoter that preferentially drives the expression of the target genes (i.e. the light chain and heavy chain of an anti-TDP-43 antibody) in cells and tissues of the CNS, preferably the human CNS. Examples of CNS-specific promoters are CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), GfaABCID (modified GFAP promoter), ATP1A2 (Na, K ATPase a2 promoter), CLDN5 (Claudin 5 promoter), ADRB2 (Adrenoceptor beta 2 promoter), TNFRSF6B (TNF receptor superfamily member 6b promoter), PDYN (prodynorphin promoter), GH1 (Human growth hormone promoter), OP ALIN (Opalin promoter), SYN1 (Synapsin 1 promoter), CAMK2A (Calcium / Calmodulin Dependent Protein Kinase II alpha promoter), NEFH (neurofilament heavy polypeptide promoter), NEUROD6 (neuronal differentiation factor 6 promoter), OLIG2 (oligodendrocyte transcription factor 2 promoter), CMVe-OLIG2 (CMV enhancer fused to OLIG2 promoter). For example, the promoter may preferentially drive expression in astrocytes. CMVe-GFAP is an astrocyte selective promoter.

[0089] In some embodiments, the AAV vector comprises an expression cassette comprising a promoter, wherein the promoter is a CMV (cytomegalovirus promoter), EF1A (Human Eukaryotic translation elongation factor 1 alpha 1), CAG (CMV early enhancer fused to modified chicken P-actin promoter), CBA (CMV early enhancer fused to chicken P-actin promoter and SV40 intron), CBh (CMV early enhancer fused to modified chicken P-actin promoter), SV40 (Simian virus 40 enhancer / early promoter), CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), GfaABCID (modified GFAP promoter), ATP1A2 (Na, K ATPase a2 promoter), CLDN5 (Claudin 5 promoter), ADRB2 (Adrenoceptor beta 2 promoter), TNFRSF6B (TNF receptor superfamily member 6b promoter), PDYN (prodynorphin promoter), GH1 (Human growth hormone promoter), OPALIN (Opalin promoter), SYN1 (Synapsin 1 promoter), CAMK2A (Calcium / Calmodulin Dependent Protein Kinase II alpha promoter), NEFH (neurofilament heavy polypeptide promoter), NEUROD6 (neuronal differentiation factor 6 promoter), OLIG2 (oligodendrocyte transcription factor 2 promoter) or a CMV early enhancer fused to either GFAP, ATP1A2, CLDN5, ADRB2, TNFRSF6B, PDYN, GH1, OPALIN, SYN1, CAMK2A, NEFH, NEUROD6 or OLIG2 promoter. In an embodiment, the promoter is a CMV, CBh, CBA, CAG, CMVe-GFAP, GFAP, CMVe-OLIG2, OLIG2 or SYN1 promoter. In a further embodiment, the promoter is a CBh, CMVe-GFAP, CMVe-OLIG2 or CBA promoter. In a preferred embodiment, the promoter is a CBh or CMVe-GFAP promoter.

[0090] In an embodiment, the promoter comprises the nucleic acid sequence of SEQ ID NO: 30. In an embodiment, the promoter comprises the nucleic acid sequence of SEQ ID NO: 31. In an embodiment, the promoter comprises the nucleic acid sequence of SEQ ID NO: 32. In an embodiment, the promoter comprises the nucleic acid sequence of SEQ ID NO: 42. Variants of these promoters may be utilised that drive expression to a comparable level. For example, variants may share at least 85%, 90%, 95%, 97%, 98% or 99% identity with SEQ ID NO: 30, 31, 32 or 42 respectively.

[0091] As described elsewhere herein, the AAV vector of the invention comprises an expression cassette, wherein the expression cassette encodes an internal ribosome entry site (IRES). In one embodiment, the IRES is derived from encephalomyocarditis virus. In a further embodiment, the IRES comprises the nucleic acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34. Variants of these IRESs may be utilised that facilitate expression to a comparable level. For example, variants may share at least 85%, 90%, 95%, 97%, 98% or 99% identity with SEQ ID NO: 33 or 34 respectively. In some embodiments, the AAV vector comprises an expression cassette comprising the light chain and the heavy chain of an anti-TDP-43 antibody, wherein the light chain of the antibody comprises a secretion peptide and / or the heavy chain of the antibody comprises a secretion peptide, wherein the secretion peptide is a mouse IgG kappa chain secretion peptide, a mouse heavy chain secretion peptide (in particular a mouse immunoglobulin heavy chain secretion peptide), a human growth hormone 1 secretion peptide, a human heavy chain secretion peptide (in particular a human immunoglobulin heavy chain secretion peptide), a human light chain secretion peptide (in particular a human immunoglobulin light chain secretion peptide), a human Oncostatin M (OSM) secretion peptide, a Vesicular stomatitis virus G protein (VSV- G) secretion peptide, a basement-membrane protein 40 (BM40) secretion peptide, a Secrecon secretion peptide, a CD33 secretion peptide, a tissue plasminogen activator (tPA) secretion peptide, a Human Chymotrypsinogen secretion peptide, a Human trypsinogen-2 secretion peptide, a Human IL-2 secretion peptide, a Gaussia luciferase secretion peptide, an Human serum albumin secretion peptide, an Influenza Haemagglutinin secretion peptide, a Human insulin secretion peptide or a Silkworm Fibroin LC secretion peptide. In a preferred embodiment, the secretion peptide is a mouse IgG kappa chain secretion peptide, a mouse heavy chain secretion peptide (in particular a mouse immunoglobulin heavy chain secretion peptide), a human growth hormone 1 secretion peptide, a human heavy chain secretion peptide (in particular a human immunoglobulin heavy chain secretion peptide) or a human light chain secretion peptide (in particular a human immunoglobulin light chain secretion peptide). In a more preferred embodiment, the secretion peptide is a mouse IgG kappa chain secretion peptide or a human growth hormone 1 secretion peptide. In an embodiment, the secretion peptide comprises the amino acid sequence of SEQ ID NO: 40 or 41. In an embodiment, the secretion peptide comprises the amino acid sequence of SEQ ID NO: 40. In a further embodiment, the secretion peptide comprises the amino acid sequence of SEQ ID NO: 41. Variants of these secretion peptide may be utilised that drive secretion to a comparable level. For example, variants may share at least 80%, 85%, 90% or 95% identity with SEQ ID NO: 40 or 41. In an embodiment, the secretion peptide is at the N-terminus of the light chain of the antibody and / or at the N-terminus of the heavy chain of the antibody. In an embodiment, the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide. In an embodiment, the secretion peptide of the light chain of the antibody and the secretion peptide of the heavy chain of the antibody are the same. In some embodiments, the AAV vector comprises an expression cassette encoding a regulatory element, wherein the regulatory element comprises a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) and / or at least one miR binding site. In some embodiments, the AAV vector comprises an expression cassette encoding a regulatory element, wherein the regulatory element consists essentially of or consists of a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) and / or at least one miR binding site.

[0092] In an embodiment, the regulatory element comprises a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE). In an embodiment, the regulatory element essentially consists of or consists of a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE). In an embodiment, the WPRE comprises the nucleic acid sequence of SEQ ID NO: 35. Variants of this WPRE may be utilised that facilitate expression to a comparable level. For example, variants may share at least 85%, 90%, 95%, 97%, 98% or 99% identity with SEQ ID NO: 35.

[0093] In another embodiment, the regulatory element comprises at least one miR binding site. In a further embodiment, the at least one miR binding site is a miR-1 binding site, miR-122 binding site, miR- 142 binding site or miR-183 binding site. In yet a further embodiment, the at least one miR binding site comprises the nucleic acid sequence of SEQ ID NO: 36, 37, 38 or 39. In a preferred embodiment, the regulatory element comprises at least one miR-122 binding site. In a further preferred embodiment, the regulatory element comprises a sequence encoding at least three miR-122 binding sites. In an embodiment, the miR binding site comprises the nucleic acid sequence of SEQ ID NO: 36 or a miR binding site having at least 80%, 85%, 90% or 95% identity therewith. In an embodiment, the miR binding site comprises the nucleic acid sequence of SEQ ID NO: 36.

[0094] In a further embodiment, the expression cassette further comprises a polyA element. In an embodiment, the polyA element is at the 3’ end of the expression cassette.

[0095] In some embodiments, the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody , and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0096] In further embodiments, the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CMVe-GFAP promoter, (ii) the light chain of an antibody , (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP- 43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17. In further embodiments, the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CMVe-0LIG2 promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP- 43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0097] In other embodiments, the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) three miR-122 binding sites, wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0098] In further embodiments, the AAV vector comprises an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBA promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0099] In some embodiments, the AAV vector is a wild type AAV vector. In some embodiments, the AAV vector is an engineered AAV vector. In an embodiment, the AAV vector is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh.8, AAVrh.9, AAVrh.10, AAVrh. l l, AAVrh.12, AAVrh.13, AAVrh.14, AAVrh.15, AAVrh.16, AAVrh.17 or AAV.Hu68. In another embodiment, the AAV vector is an engineered AAV1, engineered AAV2, an engineered AAV3, an engineered AAV4, an engineered AAV5, an engineered AAV6, an engineered AAV7, an engineered AAV8, an engineered AAV9, an engineered AAV10, an engineered AAV11, an engineered AAV12, an engineered AAVrh.8, an engineered AAVrh.9, engineered AAVrh. lO, an engineered AAVrh. l l, an engineered AAVrh.12, an engineered AAVrh.13, an engineered AAVrh.14, an engineered AAVrh.15, an engineered AAVrh.16, an engineered AAVrh.17 or an engineered AAV.Hu68. In an embodiment, the AAV vector is an AAV2-BR1, AAV-S, AAV-F, AAV.PHP.eB, AAV9.PHP.V1, AAV1RX, AAV1R6, AAV1R7, AAV.CAP-B10, AAV.CAP- B22, AAV.CAP-Mac, AAV VCAP-103, AAV bCap 1, V-CAP102, AAV-CGN1, AAV- STRV5, AAV-PAL2, AAV-MDV1A, AAV-MDV1B, AAV-MaCPNSl, AAV-MaCPNS2 or AAV VCAP-100. In an embodiment, the AAV vector is an AAV9, AAVrh.lO, AAV.PHP.eB or AAV2-BR1. In a preferred embodiment, the AAV vector is an AAV9, AAVrh.lO or AAV.PHP.eB.

[0100] As noted above, the AAV vector of the invention comprises an expression cassette that encodes the heavy chain and light chain of an anti-TDP-43 antibody.

[0101] The anti-TDP-43 antibody as defined herein: (i) binds to an epitope within amino acids 397- 411 of human TDP-43 (SEQ ID NO: 1); (ii) binds physiologically functional and pathological / misfolded TDP-43; (iii) inhibits TDP-43 aggregation; and / or (iv) reduces the level of phosphorylated TDP-43. In an embodiment, the anti-TDP-43 antibody binds TDP-43 with an affinity constant (KD) of <150 pM, preferably of <100 pM. Reference can be made to Example 7 for methods of measuring the affinity constant (KD) of an antibody.

[0102] In an embodiment, the anti-TDP-43 antibody as defined herein, binds misfolded aggregated TDP-43 and non-aggregated physiological TDP-43. In an embodiment, the anti-TDP-43 antibody binds to monomeric and / or oligomeric and / or aggregated and / or post-translationally modified and / or truncated TDP-43, preferably human TDP-43. In an embodiment, the anti- TDP-43 antibody binds to misfolded aggregated human TDP-43 and non-aggregated physiological human TDP-43.

[0103] In an embodiment, the anti-TDP-43 antibody exhibits one or more, up to all of the following characteristics: a. inhibits the aggregation of TDP-43 protein or fragments thereof, b. blocks TDP-43 cell-to-cell propagation; c. disaggregates TDP-43 aggregates; d. blocks TDP-43 seeding; e. neutralizes seeding-competent TDP-43; f. blocks TDP-43 spreading; and g. potentiates TDP-43 clearance.

[0104] In an embodiment, the anti-TDP-43 antibody neutralizes seeding-competent TDP-43. In an embodiment, the anti-TDP-43 antibody potentiates TDP-43 clearance. In an embodiment, the anti-TDP-43 antibody reduces TDP-43 pathology in vivo. In an embodiment, the anti-TDP-43 antibody reduces levels of misfolded aggregated TDP-43 and / or phosphorylated TDP-43 in vivo.

[0105] In some embodiments, the anti-TDP-43 antibody is selected from the group consisting of a full antibody (immunoglobulin, like an IgGl, an IgG2, an IgAl, an IgGA2, an IgG3, an IgG4, an IgA, an IgM, an IgD or an IgE), Fab-, Fv-, Fab’-, F(ab’)2- fragment, a humanized antibody, a chimeric antibody, a CDR-grafted antibody, a fully human antibody, an antibody-fusion protein or a synthetic antibody. In some embodiments, the antibody is a Fab, IgGl, IgG2, IgG3 or IgG4. In some embodiments, the anti-TDP-43 antibody is a chimeric or humanized antibody. In a preferred embodiment, the antibody is a Fab, IgGl or IgG4. In a more preferred embodiment, the antibody is an IgGl or IgG4. In an embodiment, the anti-TDP-43 antibody comprises an Fc mutation. In a preferred embodiment, the Fc mutation is an S228P mutation.

[0106] In an aspect of the invention, there is provided an AAV9 vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’: (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0107] In an embodiment, the AAV9 vector is administered by intracistemal (intraci sterna magna) or intracerebroventricular route.

[0108] In an aspect of the invention, there is provided an AAV9 vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’: (i) a CMVe-GFAP promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0109] In an embodiment, the AAV9 vector is administered by intracistemal (intraci sterna magna) route.

[0110] In an aspect of the invention, there is provided an AAV9 vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’: (i) a CMVe-0LIG2 promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0111] In an embodiment, the AAV9 vector is administered by intracistemal (intraci sterna magna) route.

[0112] In an aspect of the invention, there is provided an AAVrh.10 vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’: (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0113] In an embodiment, the AAVrh.10 vector is administered by intracerebroventricular route.

[0114] In an aspect of the invention, there is provided an AAV.PHP.eB vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0115] In an embodiment, the AAV.PHP.eB vector is administered by intravenous route.

[0116] In an aspect of the invention, there is provided an AAV.PHP.eB vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) three miR-122 binding sites, wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0117] In an embodiment, the AAV.PHP.eB vector is administered by intravenous route.

[0118] In an aspect of the invention, there is provided an AAV.PHP.eB vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBA promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17. In an embodiment, the AAV.PHP.eB vector is administered by intravenous route.

[0119] As noted above, the AAV vector of the invention comprises an expression cassette that encodes the heavy chain and light chain of an anti-TDP-43 antibody.

[0120] In an embodiment, the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17. In a further embodiment, the anti-TDP-43 antibody is a murine or chimeric antibody. In yet a further embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 or a VH having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 14 or a VL having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14. In still a further embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 and a VL comprising the amino acid sequence of SEQ ID NO: 14.

[0121] In an embodiment, the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27. In a further embodiment, the anti-TDP-43 antibody is a humanized antibody. In yet a further embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 or a VH having at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a VL comprising the amino acid sequence of SEQ ID NO: 24 or a VL having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24. In still a further embodiment, the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 and a VL comprising the amino acid sequence of SEQ ID NO: 24. Compositions and methods

[0122] The invention also relates to pharmaceutical compositions comprising an AAV vector of the invention as described herein and a pharmaceutically acceptable carrier and / or excipient and / or diluent.

[0123] In another aspect of the invention, there is provided an AAV vector or pharmaceutical composition (as described herein) for use as a medicament. In further aspects, an AAV vector or pharmaceutical composition (as described herein) for use in a method of treatment is provided. In yet a further aspect, there is provided an AAV vector or pharmaceutical composition (as described herein) for use in a method of reducing the level of phosphorylated TDP-43 in a subject. In an embodiment, the level of phosphorylated TDP-43 is reduced in the hippocampus of the subject.

[0124] In a further aspect, the invention provides an AAV vector or a pharmaceutical composition (as described herein) for use in prevention, alleviation or treatment of a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy. In an embodiment, the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy is Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’ s disease (PD).

[0125] In an embodiment, Frontotemporal dementia (FTD) is selected from sporadic Frontotemporal dementia, familial Frontotemporal dementia with motor-neuron disease (MND), familial Frontotemporal dementia without motor-neuron disease (MND), Frontotemporal dementia with progranulin (GRN) mutation, Frontotemporal dementia with C9orf72 mutations, Frontotemporal dementia linked to chromosome 9p, Frontotemporal dementia with TARDBP mutation, Frontotemporal dementia with valosin-containing protein (VCP) mutation, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), Argyrophilic grain disease, semantic variant Primary Progressive Aphasia (svPPA), behavioural variant FTD (bvFTD) and nonfluent variant Primary Progressive Aphasia (nfvPPA). In an embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation, Amyotrophic lateral sclerosis with C9orf72 mutation, Amyotrophic lateral sclerosis with CSL5 mutation, Amyotrophic lateral sclerosis with ALS2 mutation, Amyotrophic lateral sclerosis with ANXA1 1 mutation, Amyotrophic lateral sclerosis with ATXN2 mutation, Amyotrophic lateral sclerosis with ATXN3 mutation, Amyotrophic lateral sclerosis with C21orf2 mutation, Amyotrophic lateral sclerosis with CAV1 mutation, Amyotrophic lateral sclerosis with CCNF mutation, Amyotrophic lateral sclerosis with CHCHD10 mutation, Amyotrophic lateral sclerosis with CHMP2B mutation, Amyotrophic lateral sclerosis with CHRNA3 mutation, Amyotrophic lateral sclerosis with DAO mutation, Amyotrophic lateral sclerosis with DCTN1 mutation, Amyotrophic lateral sclerosis with DNAJC7 mutation, Amyotrophic lateral sclerosis with ELP3 mutation, Amyotrophic lateral sclerosis with ERBB4 mutation, Amyotrophic lateral sclerosis with EWSR1 mutation, Amyotrophic lateral sclerosis with FIG4 mutation, Amyotrophic lateral sclerosis with GLE1 mutation, Amyotrophic lateral sclerosis with GLT8D1 mutation, Amyotrophic lateral sclerosis with hnRNPAl mutation, Amyotrophic lateral sclerosis with hnRNPA2Bl mutation, Amyotrophic lateral sclerosis with KANK1 mutation, Amyotrophic lateral sclerosis with KIF5A mutation, Amyotrophic lateral sclerosis with LGALSL mutation, Amyotrophic lateral sclerosis with MATR3 mutation, Amyotrophic lateral sclerosis with MOBP mutation, Amyotrophic lateral sclerosis with NEFH mutation, Amyotrophic lateral sclerosis with NEKl mutation, Amyotrophic lateral sclerosis with NIPAl mutation, Amyotrophic lateral sclerosis with OPTN mutation, Amyotrophic lateral sclerosis with PARK9 mutation, Amyotrophic lateral sclerosis with PFN1 mutation, Amyotrophic lateral sclerosis with P0N1 mutation, Amyotrophic lateral sclerosis with P0N2 mutation, Amyotrophic lateral sclerosis with P0N3 mutation, Amyotrophic lateral sclerosis with PRPH mutation, Amyotrophic lateral sclerosis with SARM1 mutation, Amyotrophic lateral sclerosis with SCFD1 mutation, Amyotrophic lateral sclerosis with SETX mutation, Amyotrophic lateral sclerosis with SIGMAR1 mutation, Amyotrophic lateral sclerosis with SPG11 mutation, Amyotrophic lateral sclerosis with SPTLC1 mutation, Amyotrophic lateral sclerosis with SQSTM1 mutation, Amyotrophic lateral sclerosis with TAF15 mutation, Amyotrophic lateral sclerosis with TBK1 mutation, Amyotrophic lateral sclerosis with TIA1 mutation, Amyotrophic lateral sclerosis with TUBA4A mutation, Amyotrophic lateral sclerosis with UBQLN2 mutation, Amyotrophic lateral sclerosis with VAPB mutation, Amyotrophic lateral sclerosis with VCP mutation, Amyotrophic lateral sclerosis with WDR7 mutation and Amyotrophic lateral sclerosis with ANG mutation. In a preferred embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation and Amyotrophic lateral sclerosis with C9orf72 mutation.

[0126] In an embodiment, Alzheimer’s disease (AD) is selected from sporadic forms of Alzheimer’s disease and familial forms of Alzheimer’s disease.

[0127] In an embodiment, the Polyglutamine disease is selected from Huntington’s disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado Joseph Disease).

[0128] In an embodiment, the Myopathy is selected from sporadic inclusion body myositis and Inclusion body myopathy caused by mutation(s) in valosin-containing protein (VCP) and associated with Paget’s disease of the bone (PDB) and Frontotemporal dementia (FTD) abbreviated IBMPFD, Oculo-pharyngeal muscular dystrophy with rimmed vacuoles, Myofibrillar myopathy with mutation(s) in the myotilin (MYOT) gene and Myofibrillar myopathy with mutation(s) in the desmin (DES) gene.

[0129] In a preferred embodiment, the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), or limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0130] In a preferred embodiment, the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Alzheimer’s disease (AD). In another preferred embodiment, the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD). In another preferred embodiment, the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

[0131] A “subject” or an "individual" according to any of the embodiments of the invention may be an animal, a mammal, and is preferably a human. In another aspect, the invention provides a method of retaining or increasing cognitive memory capacity or slowing memory loss in a subject with a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, comprising administering an AAV vector or pharmaceutical composition (as described herein) to the subject. In yet another aspect, the invention provides a method of reducing the level of aggregated TDP-43 and / or phosphorylated TDP-43 in a subject, comprising administering an AAV vector or pharmaceutical composition (as described herein) to the subject. In an embodiment, administration of the AAV vector results in a reduction of the level of phosphorylated TDP-43 in a subject. In an embodiment, the level of aggregated TDP-43 and / or phosphorylated TDP-43 is reduced in the hippocampus of the subject.

[0132] In an embodiment, the provided methods further comprise administering at least one additional therapeutic. In another embodiment, the additional therapeutic agent targets alpha-synuclein, BACE1, Tau, beta-amyloid, TDP-43 or a neuroinflammation protein.

[0133] In another aspect of the invention, there is provided a method of inducing production of an anti-TDP-43 antibody in a subject, comprising administering an AAV vector of the invention (as described herein) to the subject. In yet another aspect of the invention, there is provided a method of inducing a sustained anti-TDP-43 antibody expression in a subject, comprising administering an AAV vector of the invention (as described herein) to the subject. Sustained expression generally refers to expression maintained over a period of time above a certain threshold level. The level may be sustained in the blood and / or CSF of the subject. Reference may be made to Example 4 as an example of sustained anti-TDP-43 antibody expression in a subject. The level of antibody expression may increase over a short period of time, typically between 1 and 30 days, and then reach a level that is sustained for an extended period of time.

[0134] In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the blood plasma or serum of the subject from 0.1 pg / mL to 5000 pg / mL after the administration.

[0135] In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the blood plasma or serum of the subject from 1 pg / mL to 1000 pg / mL after the administration. In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the blood plasma or serum of the subject from 1 pg / mL to 400 pg / mL after the administration.

[0136] In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the blood plasma or serum of the subject from 50 pg / mL to 250 pg / mL after the administration.

[0137] In an embodiment, the blood plasma or serum level of the anti-TDP-43 antibody is sustained at a concentration of at least 50 pg / mL, at least 100 pg / mL, or at least 200 pg / mL for a period of at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 16 weeks after the administration.

[0138] In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the CSF of the subject from 1 ng / mL to 1000 ng / mL after the administration.

[0139] In an embodiment, the provided methods achieve a concentration of the anti-TDP-43 antibody in the CSF of the subject from 50 ng / mL to 500 ng / mL after the administration.

[0140] In an embodiment, the CSF level of the anti-TDP-43 antibody is sustained at a concentration of at least 50 ng / mL, at least 100 ng / mL, or at least 200 ng / mL for a period of at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 16 weeks after the administration.

[0141] In an embodiment, the sustained concentration of anti-TDP-43 antibody is achieved after a single administration. Reference can be made to Examples 4 and 5 for methods of measuring functional antibody concentration in blood plasma or serum.

[0142] As demonstrated herein, the vectorized delivery of the anti-TDP-43 antibodies results in enhanced antibody exposure in the CNS (as compared with non-vectorized antibody delivery). The vectorized delivery of the anti-TDP-43 antibodies results in expression of the antibodies in the parenchyma. In some embodiments, when the AAV vector is administered to a subject, in particular a human subject, the anti-TDP-43 antibody is expressed in multiple brain regions. The brain regions where anti-TDP-43 antibody is expressed may depend on the promoter included in the AAV vector. The multiple brain regions may be selected from two or more of the isocortex, hippocampus and cerebellum.

[0143] In some embodiments, when the AAV vector is administered to a subject, in particular a human subject, the anti-TDP-43 antibody is expressed with a mean CSF to serum or CSF to plasma ratio of greater than 0.1%, 0.2% or more, 0.5% or more, 1% or more, 2% or more, 3% or more, 4% or more, 4.5% or more, or 5% or more. The CSF to serum ratio (CSF-to-serum ratio) or CSF to plasma ratio (CSF-to-plasma ratio) can be determined by any appropriate means. Typically, the ratio is measured by quantifying antibody titers in both sample types and then using the quantified values to derive the ratio. Antibody titers may be calculated using any suitable method, such as by an ELISA (see Examples 4 and 5 herein). Reference may be made to Examples 4 and 5 where CSF to serum or CSF to plasma ratios were calculated.

[0144] All embodiments described herein relating to AAV vectors or pharmaceutical compositions, or methods and medical uses according to the preceding aspects of the invention are equally applicable to these further aspects of the invention.

[0145] In some embodiments, the AAV vector is administered by parenteral route. In an embodiment, the AAV vector is administered by intravenous, intraperitoneal, intranasal, intravitreous, subcutaneous, intramuscular, intrathecal, intraci sternal (intraci sterna magna), intraparenchymal, intrastriatal or intracerebroventricular route. In a preferred embodiment, the AAV vector is administered by intravenous, intrathecal, intracerebroventricular, intraci sternal (intraci sterna magna), intrastriatal or intraparenchymal route.

[0146] In some embodiments, the AAV vector administration comprises delivery of from 1 • 1010vector genomes (vg) / kg to 5 - 1014vg / kg of the AAV vector to the subject per dose.

[0147] In another aspect of the invention, there is provided a method for producing an AAV vector, comprising: (i) providing a host cell comprising the viral genome of the AAV vector (as described herein); (ii) incubating the host cell under conditions suitable to enclose the viral genome in an AAV capsid, thereby making the AAV vector. All embodiments described herein relating to AAV vectors and pharmaceutical compositions according to the preceding aspects of the invention are equally applicable to this further aspect of the invention. Sequence Listing

[0148] Table 1: Amino acid sequences Table 2: Amino acid sequence of the heavy and light chain variable regions (VH and VL) and their CDRs of anti-TDP-43 antibody

[0149] BRIEF DESCRIPTION OF THE FIGURES

[0150] Figure 1. Graphical representation of the ACI-7069-633B12-Abl antibody titer in A) serum, B) brain tissue and C) cerebrospinal fluid (CSF) after antibody i.p. injections (day 0 and 24) or single vectorized delivery by i.v. or i.c.m. injection (day 0). The control is an AAV2-BR1 expressing the luciferase enzyme under the control of a CBh promoter. The data are represented as mean ± standard deviation (SD). Figure 2. IHC images of mice brain sections staining using an anti-human Fc antibody to detect ACI-7069-633B12-Abl antibody. A) Brain section of a mouse i.v. injected with a control AAV2-BR1 vector expressing luciferase. B) Brain section of a mouse i.p. injected with ACI- 7069-633B12-Abl antibody. C) Brain section of a mouse i.v. injected with an AAV2-BR1 vector expressing ACI-7069-633B12-Abl antibody under a CBh promoter. D) Brain section of a mouse i.c.m. injected with an AAV9 vector expressing ACI-7069-633B12-Abl antibody under a CBh promoter.

[0151] Figure 3. IF images of mice brain sections staining using an anti-human kappa light chain antibody to detect ACI-7069-633B12-Abl antibody. Left panel shows brain regions of mice i.p. injected with 7069-633B12-Abl antibody and arrows indicating the presence of staining in brain blood vessels. Right panel shows brain regions of mice i.c.m. injected with an AAV9 vector expressing ACI-7069-633B12-Abl antibody under a CBh promoter and arrows indicating the presence of staining in brain cells.

[0152] Figure 4. Graphical representation of the evolution of ACI-7069-633B12-Abl antibody titers in serum or plasma after a 30 mg / kg antibody i.v. injection or vectorized mAb delivery by i.c.m. injection on day 0. Dashed line represents assumption of antibody titers after study endpoint (day 25 for vectorized delivery, day 28 for mAb injection).

[0153] Figure 5. Schematic representation of the study timeline.

[0154] Figure 6. Graphical representation of ACI-7069-633B12-Abl antibody titers in A) serum and B) CSF 120 days after a single i.c.m. injection of an AAV9 vector expressing ACI-7069- 633B12-Abl antibody under a CBh or GFAP promoter. The data are represented as mean ± SD. P-values significance: *<0.05; **<0.01; ***<0.001; ****<0.0001.

[0155] Figure 7. Graphical representation of the relationship between ACI-7069-633B12-Abl antibody titers in serum and CSF 120 days after a single i.c.m. injection of an AAV9 vector expressing ACI-7069-633B 12- Ab 1 antibody under a CBh or GFAP promoter. Dashed line with crosses represents the theoretical 0.1 % CSF-to-serum ratio expected for antibodies after administration in periphery. Linear regression curve fit is shown for AAV9-CBh-mAb (light grey) and AAV9-GFAP-mAb (dark grey).

[0156] Figure 8. IF images of phosphoTDP-43, NeuN and Ibal staining on brain section of TDP-43 proteinopathy mice model treated by ACI-7069-633B12-Abl antibody vectorized delivery. Group 1 : Control. Group 2: AAV9-CBh-mAb. Group 3: AAV9-GFAP-mAb.

[0157] Figure 9. Graphical representation of pTDP-43 quantification (normalized by NeuN) in hippocampus of TDP-43 proteinopathy mice model treated by ACI-7069-633B12-Abl antibody vectorized delivery. The data are represented as mean ± SD. P-values significance: *<0.05; **<0.01; ***<0.001; ****<0.0001.

[0158] Figure 10. Schematic representation of the study timeline.

[0159] Figure 11. Graphical representation of ACI-7069-633B12-Abl or ACI-7069-633B12- Abl_H33L27 antibody titers in serum (black left histogram and white dot) and CSF (grey right histogram and black dot) 28 days after vectorized delivery using different AAV constructs and administration routes (i.c.m.: intracisterna magna injection; i.c.v.: intracerebroventricular injection; i.v.: intravenous injection). The data are represented as mean ± SD.

[0160] Figure 12. Graphical representation of ACI-7069-633B12-Abl antibody titer in serum after a 60 mg / kg antibody i.p. injection (triangles) or vectorized mAb delivery by i.c.v. injection (circles) up to 28 days post administration. The curves illustrate the assumed evolution of the antibody titer in the serum.

[0161] Figure 13. IF images of FTLD-FTD patients brain sections staining using an anti- phosphoTDP-43 antibody and ACI-7069-633B12-Abl mAb (first line) or vectorized mAbs (second and third lines). Scale bar: 10 pm.

[0162] Figure 14. A) TDP-43 aggregation kinetic measured by absorbance at 600 nm in the presence of ACI-7069-633B12-Abl mAb, vectorized mAb (VmAb) or isotype antibody control with or without TEV protease. B) Graphical representation of the % of TDP-43 aggregation in the presence of ACI-7069-633B12-Abl mAb, vectorized mAb (VmAb) or isotype antibody control at 24h. Data are represented as mean ± SD. C) Immunodetection of TDP-43 and TDP- 43-MBP on JESS blot following capillary electrophoresis.

[0163] Figure 15. Graphical representation of A) murine ACI-7069-633B12-Abl and B) chimeric ACI-7069-633B12-Abl vectorized mAb (VmAb) titers in the cell culture media of rat primary cells 7 days post transduction using an AAV9 expressing the VmAb under the control of a CMV, OLIG2, GFAP or CBh. Data are represented as mean ± SD. P-values significance: *p< 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

[0164] Figure 16. IF images of immunolabeled rat primary brain cells expressing murine ACI-7069- 633B12-Abl VmAb after transduction with AAV9 using CMV, OLIG2, GFAP or CBh promoters. Cells were immunostained for anti-mouse IgG2a to detect the VmAb expression, anti-MAP2 to identify neuronal cells and anti-GFAP to identify astrocytes. White arrows indicate the co-localization of the IgG2a and GFAP signal indicating VmAb expression in astrocytes or the co-localization of IgG2a and MAP2 indicating VmAb expression in neurons. Scale bar = 100 pm. Figure 17. Graphical representation of murine ACI-7069-633B12-Abl and chimeric ACI- 7069-633B12-Abl vectorized mAb (VmAb) titers in A) serum and B) CSF of mice 28 days post administration of an AAV9 expressing the VmAb under the control of a CMV, 0LIG2, GFAP or CBh. Data are represented as mean ± SD. P-values significance: *p < 0.05, **p < 0.01, ***p < 0.001 and **** / ? < 0.0001.

[0165] Figure 18. Graphical representation of SH-SY5Y cells mRNA levels 7 days post treatment with TARDBP shRNA, chimeric ACI-7069-633B12-Abl mAb or chimeric ACI-7069- 633B12-Abl VmAb. mRNA levels are reported as 2'AACt(Log 2 scale, y-axis) from triplicate averages.

[0166] EXAMPLES

[0167] Example 1. Aim of study and vector constructs for ACI-7069-633B12-Abl and ACI- 7069-633B12-Abl_H33L27 antibody delivery

[0168] The aim of the studies described in the following Examples was to evaluate the suitability and efficacy of a vectorized delivery approach of ACI-7069-633B12-Abl anti-TDP-43 antibody (described in WO2020 / 234473) and its humanized version ACI-7069-633B12-Abl_H33L27 (antibody described in WO2023 / 156549) for the treatment of diseases, disorders or abnormalities associated with TDP-43.

[0169] Several AAV vectors were generated comprising an expression cassette encoding the genes of the light chain and the heavy chain of the ACI-7069-633B12-Abl or ACL7069-633B12- Abl_H33L27 antibody separated by an internal ribosome entry site (IRES) allowing expression of the full-length antibody under the control of a ubiquitous or CNS-specific promoter. The antibody light chain and the antibody heavy chain were preceded by a mouse IgG kappa chain secretion peptide. The expression cassette further comprised a WPRE (except when explicitly indicated otherwise) and a poly-adenylation signal (Poly A). Table 4 details the expression cassette components and AAV vectors used in the experiments.

[0170] Table 4. Summary of expression cassette components and AAV vectors

[0171] *The expression cassette always comprises the antibody light chain gene followed by an IRES and the antibody heavy chain gene for the full length-antibody expression, and each of the light chain and heavy chain is preceded by a mouse IgG kappa chain secretion peptide. It is referred to as the “antibody”, “mAb ” (monoclonal antibody) or “VmAb ” (vectorized antibody) in the Examples.

[0172] **CMVe-GFAP and CMVe-0LIG2 promoters were referred to as GFAP and 0LIG2 promoter in the following Examples, respectively. Example 2. ACI-7069-633B12-Abl antibody titers in serum, brain and CSF after vectorized delivery or systemic injection

[0173] The aim of this study was to ascertain the brain exposure of the anti-TDP-43 chimeric monoclonal antibody ACI-7069-633B12-Abl (murine VH and VL on human IgGl and kappa constant domain, hereinafter “mAb”) either by single vectorized antibody delivery to the brain or multiple intraperitoneal injections of antibody. Twenty C57BL / 6 mice of 10 weeks of age were injected with either a control AAV vector, an AAV vector encoding the antibody, or the antibody as listed in Table 5 (N=4 mice per group).

[0174] Table 5. Summary of test articles injected in mice

[0175] At the study endpoint on day 25, CSF collection, blood collection and serum preparation were performed followed by the animal perfusion with 40 mL PBS containing 100 U / mL of heparin, collection of the brain right hemisphere fixed in 10% neutral -buffered formalin for immunohistochemistry (H4C) and immunofluorescence staining (IF), and collection of the brain left hemisphere snap-frozen for the quantification of antibody in brain tissue. The concentration of mAb in the serum, CSF and brain tissue was quantified by ELISA using an anti-human kappa light chain antibody as capture (abl25919) and an anti-human IgG Fc (ab98624) conjugated to HRP for detection. The brain exposure to the ACI-7069-633B12-Abl anti-TDP-43 mAb was confirmed by H4C using an anti -human Fc antibody (ab 109489, Abeam) conjugated to biotin (detected by streptavidin and Horseradish Peroxidase) and Acid Blue 129 (Sigma- Aldrich) counterstaining, and by IF using the same anti-human Fc antibody detected with Cy 3 -conjugated streptavidin and DAPI counterstaining on mice brain sections.

[0176] The chimeric ACI-7069-633B12-Abl mAb titer in serum reached up to 85 pg / mL using AAV9 vectorized delivery by i.e.m. injection, while AAV2-BR1 vectorized delivery by i.v. injection was unable to produce a detectable titer (Fig. 1). In comparison, the mAb i.p. injection at day 0 and 24 resulted in an average 200 pg / mL antibody serum concentration (Fig. 1), approximately a 3-fold increase compared to the AAV9-CBh vectorized delivery by i.e.m. injection. The antibody titer in brain tissue and in CSF followed the same pattern of expression level when comparing the mAh i.p. injection, AAV9 i.c.m. injection and AAV2-BR1 i.v. injection, with the mAh i.p. injection presenting a 2.5 (brain tissue) to 4-fold (CSF) higher titer than the vectorized delivery. Of note, the CSF antibody titer of the mAb i.p. injected mice are overestimated due to a possible blood contamination in CSF during sample collection.

[0177] While achieving serum, brain tissue and CSF titers, the mAb i.p. injection resulted in limited brain exposure as the chimeric ACI-7069-633B12-Abl antibody appeared to remain in the brain blood vessels as shown by IHC (Fig. 2). On the other hand, the vectorized delivery using AAV2-BR1 by i.v. injection or AAV9 by i.c.m injection were able to transduce CNS cells and achieve superior antibody exposure in the parenchyma (Fig. 2).

[0178] The results obtained in IHC for the mAb i.p. injection and AAV9-CBh vectorized delivery were confirmed by IF (Fig. 3) in different mice brain regions. ACI-7069-633B12-Abl antibody remained in the brain blood vessel after i.p. injection, while AAV9-CBh vectorized delivery was able to achieve mAb expression in several brain regions, such as the isocortex, hippocampus and cerebellum (Fig. 3).

[0179] Example 3. ACI-7069-633B12-Abl antibody blood exposure after vectorized delivery or systemic injection

[0180] The evolution of AVV9-CBh-mAb antibody titer in serum (from Example 2) was compared to the antibody plasma titer of 30 mg / kg mAb i.v. injection up to 28 days post-administration (N=9 mice, blood sample from at least 3 mice per time point). Assuming a linear decrease of the i.v. injected mAb and stable expression of the vectorized antibody, the mAb exposure of both approaches were compared (Fig. 4) and summarized in Table 6.

[0181] Table 6. Comparison of antibody exposure between delivery method

[0182] *Day28Prefers to the AUC of AAV9-CBh-mAb for every 28-day period following the initial 28 days post i.c.m. administration (assuming sustained mAb expression from day 28 postadministration). ACI-7069-633B12-Abl delivered by a single i.c.m. injection of AAV9-CBh was able to match the mAb concentration obtained by a monthly 30 mg / kg. Noteworthy, the vectorized mAb delivery was able to match mAb concentration in serum of i.v. injection despite a delivery in the CNS (via i.c.m. injection). These results suggest a longer lasting mAb brain exposure through vectorized mAb delivery to the CNS and confirm the suitability of such an approach to obtain sustained mAb titer in the brain for the treatment of CNS diseases.

[0183] Example 4. In vivo efficacy of vectorized ACI-7069-633B12-Abl antibody delivery in a TDP-43 proteinopathy mouse model

[0184] The aim of this study was to determine the efficiency of the vectorized anti-TDP-43 chimeric monoclonal antibody ACI-7069-633B12-Abl (hereinafter “mAb”) in a TDP-43 proteinopathy mouse model described in Porta et al., 2018. The CamIIKa-hTDP-43NLSm transgenic mouse is a model of ALS / FTD, expressing cytoplasmic human TDP-43 in excitatory neurons. Intracerebral injections of biologically active pathogenic FTLD-TDP seeds from patients leads to the induction of de novo TDP-43 pathology and recapitulates TDP-43 spreading throughout the brain via cell-to-cell transmission, correlating with disease progression found in human. Briefly, CamIIKa-hTDP-43NLSm mice were kept on food comprising doxycycline from birth until 13-weeks of age to prevent human TDP-43 cytoplasmic expression. Starting from 13- weeks of age, doxycycline was removed (allowing hTDP-43 cytoplasmic expression), followed by FTLD-FTD human brain extracts injection at 14-weeks of age (Porta et al., 2018). A single i.c.m. injection of the vectorized mAb under a CBh or GFAP promoter (AAV9-CBh-mAb or AAV9-GFAP-mAb) was performed at 10-weeks of age to allow sufficient time for the mAb expression. Three months after FTLD-FTD brain extracts injection (=120 days post- AAV injection), the mAb titer in plasma and CSF (cerebrospinal fluid) was measured by ELISA and the level of phosphorylated TDP-43 (phospho-TDP-43, a known marker of TDP-43 proteinopathy; Neumann et al., 2009) was quantified in the mice hippocampus by IF. The study timeline is shown in Figure 5 and study details are summarized in Table 7, respectively.

[0185] Table 7. Study details *The dose was not adjusted to the mice weight. However, the mice presented similar weight, therefore the dose administered to each mouse was approximately 4- 10I2vg / kg.

[0186] Functional plasma and CSF mAb titers

[0187] Plasma and CSF were collected before mice euthanasia at study endpoint (one mouse per group could not be sampled resulting in N=14). The mAb titers were quantified by ELISA using 96- well plates coated with 2.5 pg / ml TDP-43 (Selvita) for capture of the ACI-7069-633B12-Abl antibody, and an anti-human IgG Fc secondary antibody (Abeam, ab98624) for detection. Noteworthy, the ELISA only quantifies antibodies that can bind TDP-43.

[0188] ACI-7069-633B12-Abl delivered by a single i.e.m. injection of AAV9-CBh reached a mean titer of 195 pg / mL in plasma and 208 ng / mL in CSF at 120 days post-injection (Fig. 6). A similar plasma titer of 212 pg / mL was obtained when ACI-7069-633B12-Abl was delivered by AAV9-GFAP. However, the mean CSF titer for AAV9-GFAP-mAb was substantially higher, reaching 400 ng / mL, potentially due to a predominant in situ antibody production using astrocyte selective GFAP promoter.

[0189] The linear relationship between plasma and CSF antibody titer is shown in Figure 7, along with the theoretical 0.1% CSF-to-plasma ratio expected (St-Amour et al., 2013; Pardridge et al., 2019) for antibodies after peripheral administration (e.g. intravenous) as a dashed line with crosses. A favourable CSF-to-plasma ratio (AAV9-CBh: 0.106%; AAV9-GFAP: 0.188%) was obtained with the vectorized delivery of ACI-7069-633B12-Abl compared to mAb systemic administration. These results confirm the suitability of a vectorized antibody delivery approach to achieve a sustained antibody expression in the CNS.

[0190] In vivo efficacy of vectorized mAb

[0191] Following terminal CSF and blood collection, mice were perfused with 20-40 mL of PBS. The brains were immediately extracted, part of the brains comprising the hippocampus dissected and immersion-fixed in formalin for 24 hours, and then transferred to 30% sucrose until sunken (one or two days). The brain tissues were stored at -80°C prior to cutting. Frozen tissue blocks were sectioned at 20 pm thickness per section and collected from three coronal levels (anterior and posterior to the FTLD-FTD human brain extract inoculation site) prior to IF staining. Triple pTDP43 / NeuN / Ibal IF staining were performed using phospho-TDP-43 Ab (Biolegend, Ref. 829901, Rat Ab), NeuN (Millipore, Ref MAB377(CH), Mouse Ab) and Ibal (Wako, Ref 019-19741, Rabbit Ab) primary antibodies, followed by anti-mouse-Cy3 (Jackson, Goat Ab), anti-Rabbit-Alexa488 (Jackson, Goat Ab) and anti-Rat-biotin (Jackson, Goat Ab) secondary antibodies. Anti-Rat-biotin antibody was detected with Streptavidin-Cy5 (Jackson) and the sections counterstained with DAPI.

[0192] The IF slides were digitized using an Axio Scan.Zl digital whole slide scanner (Carl Zeiss, Canada). The images underwent quality control (QC) review before processing and analysis. ROIs (region of interests) were delimited using a U-Net convolutional neural network trained on a dataset of manually painted tissue sections, confirmed by visual QC review and manually adjusted if needed. Quantification of staining (% area stained) was performed on each of the digitized H4C slides using the Biospective PERMITS™ software. Double co-localization of pTDP-43 and NeuN, and triple co-localization of pTDP-43, NeuN, and Ibal were calculated from the segmented images. The H4C analysis and quantification was performed in a blinded manner with respect to cohort. Any potential outliers for technical reasons were removed before un-blinding the data. Statistical analyses were performed in MATLAB. Treatment groups were compared using one-way ANOVA with Tukey’s honest significant difference for post-hoc comparisons or, for non-normally distributed clinical data, using Kruskal-Wallis.

[0193] Representative IF staining of NeuN, pTDP43 and Iba-1 in the hippocampus of the right brain hemisphere (inoculated with the FTLD-FTD human brain extract) of control (Group 1), AAV9- CBh-mAb (Group 2) and AAV-GFAP-mAb (Group 3) are shown in Figure 8. Quantification of phosphoTDP-43 level (normalized to NeuN) in the hippocampus of mice treated with AAV9-CBh-mAb and AAV9-GFAP-mAB demonstrated a significant reduction in pTDP-43 level compared to control mice (Fig. 9). An average reduction of 56% and 67% pTDP43 level was achieved 120 days after a single injection of AAV9-CBh-mAb (ubiquitous promoter) and AAV9-GFAP-mAb (astrocytes selective promoter), respectively. These results confirm the vectorized delivery of ACI-7069-633B12-Abl antibody as a suitable approach to treat TDP- 43 proteinopathy. Example 5. ACI-7069-633B12-Abl and ACI-7069-633B12-Abl_H33L27 antibody titers in CSF and serum after vectorized delivery by different AAV constructs

[0194] The aim of this study was to further investigate the brain exposure of the chimeric ACI-7069- 633B12-Abl (murine VH and VL on human IgGl and kappa constant domain, hereinafter “mAb”) and humanized anti-TDP-43 monoclonal antibody ACI-7069-633B12-Abl_H33L27

[0195] (hereinafter “hmAb”) delivered by different AAV serotypes. Seven AAV vectors were compared comprising either a single intracerebroventricular (i.c.v), intracistema magna (i.c.m) or intravenous (i.v.) injection of the vectorized mAb or hmAb under a constitutive promoter, and optionally comprising a miR-122 BS. The antibody serum and CSF titer were quantified at day 28 post-injections in C57BL / 6 mice. The study timeline is shown in Figure 10 and study details are summarized in Table 8, respectively.

[0196] Table 8. Study details

[0197] *AAV.PHP .eB-CBh-mAb-miR-122BS comprises 3 miR-122 binding sites in tandem and no WPRE.

[0198] Serum and CSF mAb titers

[0199] Serum and CSF were collected before mice euthanasia at study endpoint on day 28. Functional mAb titers were quantified by ELISA as described in Example 4 and are shown in Figure 11. Invalid ELISA quantification or potential CSF samples contaminated with blood were excluded from analysis and led to a reduction in some groups (conserving at least n=2).

[0200] AAV9-CBh-mAb i.c.m injection elicited serum and CSF antibody titers in line with previous results from Example 2 and 4, and similar to AAV9-CBh-mAb and AAVrh.10-CBh-mAb i.c.v. injection. Humanized ACI-7069-633B12-Abl_H33L27 antibody delivered by AAV9-CBh- hmAb i.c.v injection reached similar serum and CSF titers as the chimeric ACI-7069-633B12- Abl. The chimeric ACI-7069-633B12-Abl antibody delivered by the AAV.PHP.eB-CBh- mAb reached the highest CSF titer of all AAV constructs with a mean titer of 463 ng / ml, while its serum titer was in range with the AAV9 and AAVrh.10 vectors.

[0201] Interestingly, the AAV-PHP.eB-CBh-mAb comprising miR-122 binding sites (which silences selectively the transgene expression in the liver) showed a ~5-fold reduction in CSF antibody titer and a ~45-fold reduction in serum antibody titer compared to AAV.PHP.eB-CBh-mAb. Without wishing to be bound by any particular hypothesis it is understood that the antibody titer in serum is mainly driven by expression of the antibody in the liver and secretion in the blood stream, therefore the presence of miR-122BS in the expression cassette strongly reduced the antibody expression and serum titer. In turn, the decrease in serum antibody titer most likely impacted CSF antibody titer by reducing the number of antibodies crossing the BBB and reaching the brain from the circulation.

[0202] The AAV.PHP.eB-CBA-mAb also showed similar serum and CSF antibody titers to AAV- PHP.eB-CBh-mAb-miR-122BS. This result suggests that the CBA promoter is weaker than the CBh promoter, and / or preferentially promotes expression in CNS cells.

[0203] The different vectorized antibody delivery approaches were further compared by calculating their CSF-to-serum ratio (Table 9) as an indicator of which strategy favours antibody brain exposure.

[0204] Table 9. CSF-to-serum ratio All the AAV constructs tested reached a similar or favourable CSF-to-serum ratio compared to the expected theoretical ratio of 0.1% obtained from systemic mAb administration (as discussed in Example 3). Altogether, the results confirm vectorized antibody delivery as a suitable strategy for CNS immunotherapy. Additionally, the results suggest that engineered AAV capsids such as PHP.eB are appropriate candidates for vectorized delivery of antibodies to the CNS after i.v. administration, inducing favourable antibody CSF-to-serum ratio. Finally, the results also suggest that the selection of an appropriate promoter in the expression cassette can substantially impact the antibody CSF-to-serum ratio and, therefore, the antibody brain exposure.

[0205] Example 6. Pharmacokinetic of ACI-7069-633B12-Abl antibody after vectorized delivery or systemic injection

[0206] The pharmacokinetics in serum and CSF of ACI-7069-633B12-Abl antibody either delivered by AAV9-CBh-mAb i.c.v. injection (as in Example 5) or by 60 mg / kg mAb i.p. injection up to 28 days post-administration was compared. Mice injected i.p. with the mAb were sampled for blood at Ihr, 4hr, 8hr, 16hr, 1 day, 4 days, 10 days, 17 days, 22 days and 28 days post-injection, and for CSF at 1 day and 28 days post injection (N=15 mice in total, at least 3 mice per time point). The mice injected i.c.v. with AAV9-CBh-mAb were sampled for blood at -1 day, 12 days and 28 days post-injection and for CSF 28 days post-injection (N=3). Both approaches were compared (Fig. 12) and are summarized in Table 10.

[0207] Table 10. Comparison of antibody pharmacokinetic between delivery method

[0208] *Cmax in serum is at 16hr and in CSF at 24hr for mAb i.p. injection, Cmax in serum and CSF for mAb vectorized delivery is at study endpoint (28 days).

[0209] ACI-7069-633B12-Abl delivered by a single i.c.v. injection of AAV9-CBh at 4 1012vg / kg induced a lower maximum concentration (Cmax) compared to the mAb i.p. injected at 60 mg / kg. However, importantly, the vectorized delivery provided longer-lasting high antibody titer in serum (>250 pg / mL from day 12 onward) compared to the mAb i.p. injection that reached titer above 200 pg / mL only the first 1-2 days post-injection (<100 pg / mL from day 3 onward). A similar trend was observed with the CSF antibody titer.

[0210] The Area Under the Curve (AUC) of the antibody titers in serum and CSF from day 0 until day 28 (AUCo-28) post administration and for every 28-day period thereafter (AUC28P; for the vectorized delivery only) are compared in Table 11.

[0211] Table 11. Comparison of antibody titers AUC between delivery method

[0212] *AUC28prefers to the AUC of AAV9-CBh-mAb for every 28-days period following the initial 28 days post i.c.v. administration (assuming sustained mAb expression from day 28 postadministration).

[0213] **Not applicable: the AUCo-28 is not calculated due to limited data point.

[0214] The serum titer AUC28Pof the vectorized antibody showed a 7-fold higher antibody exposure in blood compared to the antibody delivered by i.p. injection (AUCo-28). A similar trend was observed with the CSF titer AUC28Pthat showed a 3-fold higher antibody exposure with the vectorized antibody compared to the systemically injected antibody (AUCo-28). These results suggest a longer lasting mAb brain exposure through vectorized mAb i.c.v. delivery and confirm the suitability of such an approach to obtain a sustained mAb concentration in the brain for the treatment of CNS diseases.

[0215] Example 7. Functional properties of vectorized ACI-7069-633B12-Abl antibody

[0216] The functional properties of murine and / or chimeric ACI-7069-633B12-Abl antibody purified from CHO cells transfected with the AAV vector expression cassette (hereinafter “VmAb”) were compared to recombinant ACI-7069-633B12-Abl antibody (hereinafter “mAb) produced from established standard methods comprising co-transfection of two plasmids: one for the expression of the antibody light chain and one for the expression of the antibody heavy chain. Production of mAb and VmAb

[0217] CHO transfections were performed using the ExpiCHO™ transient expression system (Gibco), in 24 deep well blocks, according to the supplier protocol. In brief, triplicates of 2.5 mL cells (4-5 mio / mL) were transfected each with 1 pg / mL plasmid mixed to OptiPro complexation medium supplemented by Expifectamine and grown for ~18h at 37°C. After that, cells received per well 600 pL Feed supplemented by ExpiCHO enhancer to reach a final volume of 3.1 mL. Temperature was dropped to 32°C and cells were grown until harvest.

[0218] One hundred microliter samples were taken per clone for each triplicate and centrifuged 2 min at room temperature, 3000 rpm, in microfuge tubes. Supernatants were separated into fresh tubes and titers were estimated in each supernatant triplicate using Bio-Layer Interferometry (BLI) with the Octet system and biosensor tips coated with protein A. For all measures, a standard curve was performed using the corresponding and previously purified protein. Reaction buffer was PBS supplemented by 0.1% BSA and 0.02% Tween.

[0219] The productions were combined to reach about 9 mL in a single 15 mL falcon tube. Cells were harvested by centrifugation 2 min at room temperature, 3000 rpm. Supernatants were separated and added to new 15 mL falcon tubes. 300 pL equilibrated resin in PBS was added to the supernatant. Tubes were then rotated gently for 30 min at room temperature to produce a homogenous mixture of supematant / resin and allow capture of expressed proteins. Tubes were then centrifuged at 6000 rpm, room temperature and supernatants were discarded. Resins were then washed with PBS buffer followed by centrifugation as above. Resin wash was repeated once. After that, resin-captured antibodies were resuspended in 1 mL 100 mM glycine pH 2.8 supplemented by 100 mM NaCl, transferred to a 1.5 mL microfuge tube and centrifuged 5 min at 20K rpm, 4°C. Supernatants were then separated and neutralized with 200 pL IM Tris-HCl pH 7.6. Proteins were then quantified by OD 280 nm using their coefficient of extinction.

[0220] Immunofluorescence on brain sections from FTLD-TDP type A patients

[0221] Human post-mortem tissues from FTLD-FTD type A patients were obtained from the brain banks affiliated with the University of California, San Francisco (UCSF). The brain samples were cut at 10 pm thickness, mounted on microscope slides and stored at -80°C until further use. Frozen brain sections were thawed at room temperature (RT) and fixed with 4% paraformaldehyde (PF A) at 4°C for 15 min. The sections were incubated overnight at 4°C with murine ACI-7069-633B12-Abl mAb, murine ACI-7069-633B12-Abl VmAb or chimeric ACI-7069-633B12-Abl VmAb at 1 pg / mL, and p TDP-43 (s409 / 410) antibody (Biolegend, 829901) at 1 :200. As secondary antibody, Alexa Fluor-633-labeled anti-rat and Alexa Fluor- 488-labeled anti-mouse or Cy3 anti-human diluted 1 :500 in PBS were used. Prolong Antifade Reagent with DAPI was used for counterstaining. The brain sections were dried in the dark and imaged with the Panoramic 150 Slide Scanner using the fluorescein isothiocyanate (FITC), Cy5 or Cy3 and DAPI channels.

[0222] Both the mAb and VmAbs labelled pathological phosphorylated TDP-43 inclusions and physiological nuclear TDP-43 in brain sections from FTLD-TDP type A patients (Fig. 13), confirming AAV vectorized ACI-7069-633B12-Abl antibodies are functional.

[0223] Affinity measurements to human TDP-43

[0224] Target binding affinities were measured by Surface Plasmon Resonance (SPR) performed on a Biacore 8K instrument (Cytiva). Recombinant human TDP-43 (Selvita) was immobilized on flow-cells 1 and 2 (fcl and fc2 hereinafter) of channels 1 to 8 of a Series S CM5 sensor chip (Cytiva, cat# BR100530) to reach 350 to 450 Response Unit (RU) on all eight channels. Five successive increasing concentrations of murine / chimeric ACI-7069-633B12-Abl mAb or VmAbs ranging from 1.2 to 100 nM prepared from a three 3-fold serial dilution in a phosphate buffer saline containing detergent (PBS-P+ pH 7.4, Cytiva) running buffer were injected in single-cycle kinetics on fcl and fc2. Kinetic measurements were performed with a contact time of 300 s and a dissociation time of 3600 s at a flow rate of 30 pL / min. Regeneration was performed using two consecutive injections of 10 mM glycine-HCl pH 1.7, at a flow rate of 30 pL / min for 30 s. Results obtained from single-cycle kinetics were double-referenced using the blank fcl and evaluated with the Biacore Insight evaluation software. Sensorgrams were analyzed using the 1 : 1 binding model with variable reflective index and global Rmax parameters and the kinetic constants of two replicated measures (mean ± SD) are shown in Table 12.

[0225] Table 12. ACI-7069-633B12-Abl mAb and VmAb affinity to human TDP-43

[0226] SPR binding measurements to human TDP-43 showed that the vectorized mAbs (chimeric and murine) bind the target with the same picomolar affinity as their recombinant equivalent. These results confirm that the ACI-7069-633B12-Abl antibody conserves its high affinity for TDP- 43 when produced by an AAV vector.

[0227] Inhibition of TDP-43 aggregation

[0228] Murine ACI-7069-633B 12-Abl mAb and VmAb were characterized in an in vitro functional assay relying on the aggregation property of TDP-43 (Wang et al., 2018). Human TDP-43 C- terminally fused to maltose binding protein (MBP) by a Tobacco Etch Virus (TEV) protease cleavage site was recombinantly produced in E. coh. aliquoted and stored at - 80° C. Storage buffer (20 mM Tris-Cl pH 8.0, 300 mM NaCl, 5% glycerol, 1 mM DTT) was exchanged against assay buffer (30 mM Tris, 150 mM NaCl, pH 7.4) using Centrifugal Filters and the protein concentration was determined by ultraviolet (UV) spectroscopy at 280 nm (NanoDrop). TDP- 43-MBP was diluted in assay buffer to a final concentration of 2.5 pM and mixed with 600 nM mAb, VmAb, or isotype mAb control in low binding tubes. Aggregation was induced by addition of TEV protease at a final concentration of 10 pg / mL. Aggregation was monitored by absorbance measurement at 600 nm in the center of the well every 15 min, with 5 s shaking before each measurement over 24 h in technical triplicates (Figure 14A). The samples were analyzed by immunodetection after 24 h in order to confirm TEV protease cleavage in the presence of mAbs. 4 pL samples were mixed with 1 pL 5* Mastermix and analyzed in a Separation Module using capillary electrophoresis (Jess, Proteinsimple) using ACI-7069- 633B 12-Abl labeled with DyLight680 as detection mAb at 20 pg / mL. Figure 14C confirms the proper cleavage of MBP from TDP-43 in the presence of TEV protease. For analysis, end points measured by absorbance after 24 h were normalized to isotype antibody control and % aggregated TDP-43 was calculated for each antibody.

[0229] Both the mAb and VmAb inhibited TDP-43 aggregation at similar level compared to the isotype control mAb as shown in Figure 14B. These results confirm that the ACI-7069- 633B 12-Abl antibody conserves its TDP-43 anti -aggregation properties when produced by an AAV vector. Altogether, the results confirm that the ACI-7069-633B12-Abl antibody retains its functional properties when produced by an AAV vector.

[0230] Example 8. Murine and chimeric ACI-7069-633B12-Abl antibody expression after vectorized delivery in rat primary brain cells

[0231] The expression of murine and chimeric ACI-7069-633B12-Abl antibodies was evaluated in rat primary brain cells. Both antibodies were delivered by an AAV9 vector under the control of a CMV, 0LIG2, GFAP or CBh promoter.

[0232] Briefly, rat pups at post-natal day 1 (Prague-Dawley, Charles River) were euthanized and dissected in HBSS._The brains were kept in MEM (Gibco, 12360-038) before cutting and transfer into a digestion medium (30 mM K2SO4, 90 mM Na2SO4, 5.8 mM MgC12, 0.25 mM CaC12, 1.6 mM HEPES, 0.2 mM NaOH, 0.5% phenol red, 20 U / mL Papain, 0.1 % (w / v) L- Cysteine, 0.36 % (w / v) Glucose) for 30 minutes at 37°C. Supernatant was removed and 10 mL of protease inhibitor medium was added to the tissue (30 mM K2SO4, 90 mM Na2SO4, 5.8 mM MgC12, 0.25 mM CaC12, 1.6 mM HEPES, 0.2 mMNaOH, 0.5% phenol red, 0.36 % (w / v) Glucose, 0.1 % (w / v) trypsin inhibitor (Sigma, T9253)) which was incubated for 10 minutes, centrifuged at 300 g for 2 minutes and the supernatant then discarded. Tissue was resuspended in 5 mL of trituration medium (10 % horse serum (Sigma, Hl 138), 1% L-Glutamine (Gibco, 25030-025), 0.36% (w / v) Glucose in MEM) and triturated using a serological pipette. Once all the tissue was dissolved, the tube was centrifuged at 300 g for 2 minutes, the supernatant discarded, the cells resuspended in 10 mL of adhesion medium (10% horse serum, 1% L- Glutamine, 1% Pen / Strep, 0.36% (w / v) glucose in MEM) and passed through a 70 pm cell strainer. Cells were counted and plated at 40’000 cells / well in 96-well Poly-D-Lysin-coated plates in 100 pL of adhesion medium and incubated at 37°C and 5% CO2. After 4 hours, the medium was changed to a growth medium: 1% L-Glutamine, 1% Pen / Strep, 2% B27 (Gibco, 17504-044 in Neurobasal medium (Gibco, 12348-017)). After 5 days of culture, cells were transduced with the AAV9 constructs at a MOI of 100,000 to 130’000 and the ACI-7069- 633B12-Abl VmAb titers in culture media were determined 7 days post-transduction by ELISA as described in the previous Examples.

[0233] Similar titers were observed for the murine and chimeric VmAb (Figure 15A and 15B, respectively). The CMV promoter yielded the lowest VmAb titer with 70 pg / L 7 days posttransduction. The OLIG2 and GFAP promoters significantly increased the VmAb titer by 3- fold compared to the CMV promoter while the CBh promoter significantly increased the VmAb titer by 6-fold, resulting in a yield > 400 pg / L VmAb (Figure 15A and 15B).

[0234] To determine which cell type from the rat primary brain cell cultures was producing the VmAb, immunolabeling was performed using an anti-mouse IgG2a antibody (to detect the murine VmAb), an anti-MAP2 antibody for neurons and an anti-GFAP antibody for astrocytes. CMV, CBh and GFAP promoters resulted in VmAb expression in astrocytes (Figure 16). Surprisingly, the OLIG2 promoter also resulted in astrocyte VmAb expression. CBh and OLIG2 promoters demonstrated expression in neurons as well (Figure 16).

[0235] Altogether, these results confirm the expression and secretion of ACI-7069-633B12-Abl VmAb in astrocytes and neurons after AAV9 vectorized delivery. In addition, these results demonstrate that astrocyte-selective expression can be achieved with the GFAP promoter, while the OLIG2 (CNS specific) and CBh (ubiquitous) promoters result in ACI-7069-633B12- Abl VmAb expression in astrocytes and neurons.

[0236] Example 9. Murine and chimeric ACI-7069-633B12-Abl antibody titers in serum and CSF after vectorized delivery in mouse

[0237] The expression of murine and chimeric ACI-7069-633B12-Abl antibodies was evaluated in mice. Both antibodies were delivered by an AAV9 vector under the control of a CMV, OLIG2, GFAP or CBh promoter. Briefly, C57BL / 6 WT mice were injected (i.c.m. route) with a single dose of AAV9 at 4.1012vg / kg on Day 0 and serum and CSF were collected on Day 28. Antibody titers were determined using the TDP -43 -based ELISA as described in the previous Examples.

[0238] Similar antibody serum titers were observed for the murine and chimeric VmAb (Figure 17A). The CMV promoter yielded the lowest titers, with less than 5 pg / mL VmAbs detected on Day 28 in serum. The OLIG2 promoter yielded 17 pg / mL and 26 pg / mL, and the GFAP promoter 41 pg / mL and 24 pg / mL for the murine and chimeric VmAbs, respectively. The CBh promoter produced higher VmAb titers compared to other promoters, achieving mean titers above 60 pg / mL in serum. In CSF (Figure 17B), a similar antibody titer ranking across the promoters was observed with both the GFAP and CBh promoters yielding a high VmAb expression reaching over 100 ng / mL 28 days post vector administration. Altogether, these results confirm reliable in vivo vectorized delivery of ACI-7069-633B 12-Ab 1 murine and chimeric antibodies under the control of 0LIG2, GFAP or CBh promoters. The results also confirm the suitability of the vectorized delivery approach of the antibodies described herein to produce sustained antibody titers in serum and CSF.

[0239] Example 10. ACI-7069-633B12-Abl VmAb expressing cells conserve physiological TDP-43 function

[0240] The effect of ACI-7069-633B 12-Ab 1 on TDP-43 physiological function was investigated in SH-SY5Y cells. Briefly, SH-SY5Y cells were grown at 100,000 cells per well in 24-well plates and differentiated in DMEM high glucose medium (Gibco, 11965092) supplemented with 2% fetal bovine serum (Gibco, 10500064), 1% penicillin-streptomycin (Gibco, 15140122) and 10 pM of retinoic acid (Thermo Scientific Chemicals, 044540.04). Following differentiation, SH- SY5Y cells were either transduced with an AAV6 construct (as set out in Table 13) at a MOI (multiplicity of infection) of 130,000 or treated with ACI-7069-633B 12-Ab 1 (mAb in Figure 18) spiked into the cell supernatant at a 20 pg / mL final concentration.

[0241] Table 13. AAV6 constructs transduced in SH-SY5Y cells

[0242] After 7 days of cell culture, mRNA was extracted using GeneSet RNA purification kit (Thermo Fisher Scientific, cat# K0732) and mRNA levels of TARDBP, STMN2 and POLDIP3v2 genes were analyzed by RT-qPCR and normalized with RPLP0 as housekeeping gene. The mRNA levels of truncated STMN2 or POLDIP3v2 were used as an indicator of impaired TDP-43- dependent splicing (Shiga et al., 2012; Klim et al., 2019).

[0243] AAV-delivered shRNA targeting TARDBP mRNA decreased TDP-43 levels resulting in an increase in truncated STMN2 and POLDIP3v2 expression, confirming that reduction in TDP- 43 levels impairs TDP-43 splicing function (Figure 18). Neither the transduction of AAV6- CBh-VmAb (resulting in the expression of ACI-7069-633B 12-Ab 1 VmAb) nor treatment with ACI-7069-633B12-Abl mAb changed TARDBP, truncated STMN2 or POLDIP3v2 mRNA levels (Figure 18). These results confirm that ACI-7069-633B12-Abl mAb and VmAb do not interfere with endogenous TDP-43 physiological function, supporting the safety of the therapeutic vectorized approach presented herein.

[0244] Example 11. Functional properties of vectorized ACI-7069-633B12-Abl antibody expressed in vivo

[0245] The binding affinity to human TDP-43 of the vectorized ACI-7069-633B12-Abl antibody (hereinafter “VmAb”) expressed in vivo was assessed by SPR. The VmAb was isolated from the serum of mice from the experiment described in Example 3 (serum collection 28 days postadministration of the AAV vector). The VmAb binding affinity to TDP-43 was compared to recombinant ACI-7069-633B12-Abl antibody (hereinafter “mAb”) isolated from the serum of mice from the experiment descried in Example 3 (serum collection 1 day post-administration of the mAb).

[0246] Affinity measurements to human TDP-43

[0247] Target binding affinities were measured by Surface Plasmon Resonance (SPR) performed on a Biacore 8K instrument (Cytiva). The VmAb and mAb were captured on the Biacore chip using an immobilized anti-human antibody from the Human Antibody Capture Kit, type 2 (Cytiva) via amine coupling on flow-cells 1 and 2 (fcl and fc2 hereinafter) of a Series S CM5 sensor chip (Cytiva, cat# BR100530). Mouse serum was diluted approximately 100-fold in a phosphate buffer saline containing detergent (PBS-P+ buffer at pH 7.4, Cytiva) containing 10 mg / mL non-specific binding reducer additive (NSB reducer, Cytiva) to achieve a final concentration of 3 pg / mL of ACI-7069-633B12-Abl mAb or VmAbs. The diluted serum was injected in fc 2 at a flow rate of 5 pL / min to capture in vzvo-produced VmAb and mAb at levels ranging from 221 to 308 Response Unit (RU). Single-cycle kinetic analysis was performed by injecting five successive, increasing concentrations of recombinant full-length soluble human TDP-43 (Selvita) prepared in PBS-P+ (pH 7.4) running buffer on fcl and fc2. Kinetic measurements were performed with a contact time of 300 s and a dissociation time of 900 s at a flow rate of 30 pL / min. Regeneration was achieved through an injection of 10 mM glycine- HC1 (pH 1.5) at a flow rate of 30 pL / min for 60 seconds. Results obtained from single-cycle kinetics were double-referenced using the blank fcl and evaluated with the Biacore Insight evaluation software. Sensorgrams were analysed using the 1 : 1 binding model with variable reflective index and global Rmax parameters and the kinetic constants for groups are shown in Table 14.

[0248] Table 14. ACI-7069-633B12-Abl mAb and VmAb affinity to human TDP-43

[0249] SPR binding measurements to human TDP-43 showed that the vectorized mAb (VmAb) expressed in vivo binds the target with the same picomolar affinity as the mAb. These results confirm that vectorized delivery approach described herein provides sustained titer of functional ACI-7069-633B12-Abl antibody.

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[0323] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications and patents specifically mentioned herein are incorporated by reference in their entirety for all purposes in connection with the invention.

[0324] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims. Moreover, all aspects and embodiments of the invention described herein are considered to be broadly applicable and combinable with any and all other consistent embodiments, including those taken from other aspects of the invention (including in isolation) as appropriate.

Claims

WHAT IS CLAIMED IS:

1. An adeno-associated virus (AAV) vector comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a regulatory element, wherein the light chain of the antibody comprises a secretion peptide and / or the heavy chain of the antibody comprises a secretion peptide and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

2. The AAV vector of claim 1, wherein the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL- CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

3. The AAV vector of claim 2, wherein the anti-TDP-43 antibody is a murine or chimeric antibody.

4. The AAV vector of claim 2 or 3, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 or a VH having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 14 or a VL having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14.

5. The AAV vector of any one of claims 2 to 4, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 and a VL comprising the amino acid sequence of SEQ ID NO: 14.

6. The AAV vector of claim 1, wherein the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL- CDR3 comprising the amino acid sequence of SEQ ID NO: 27.

7. The AAV vector of claim 6, wherein the anti-TDP-43 antibody is a humanized antibody.

8. The AAV vector of claim 6 or 7, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 or a VH having at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a VL comprising the amino acid sequence of SEQ ID NO: 24 or a VL having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24.

9. The AAV vector of any one of claims 6 to 8, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 and a VL comprising the amino acid sequence of SEQ ID NO: 24.

10. The AAV vector of any one of the preceding claims, wherein the promoter is a ubiquitous or CNS-specific promoter.

11. The AAV vector of any one of the preceding claims, wherein the promoter is a CMV (cytomegalovirus promoter), EFl A (Human Eukaryotic translation elongation factor 1 alpha 1), CAG (CMV early enhancer fused to modified chicken P-actin promoter), CBA (CMV early enhancer fused to chicken P-actin promoter and SV40 intron), CBh (CMV early enhancer fused to modified chicken P-actin promoter), SV40 (Simian virus 40 enhancer / early promoter), CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), GfaABCID (modified GFAP promoter), ATP1A2 (Na, K ATPase a2 promoter), CLDN5 (Claudin 5 promoter), ADRB2 (Adrenoceptor beta 2 promoter), TNFRSF6B (TNF receptor superfamily member 6b promoter), PDYN (prodynorphin promoter), GH1 (Human growth hormone promoter), OPALIN (Opalin promoter), SYN1 (Synapsin 1 promoter), CAMK2A (Calcium / Calmodulin Dependent Protein Kinase II alpha promoter), NEFH (neurofilament heavy polypeptide promoter), NEUROD6 (neuronal differentiation factor6 promoter), 0LIG2 (oligodendrocyte transcription factor 2 promoter) or a CMV early enhancer fused to either GFAP, ATP1 A2, CLDN5, ADRB2, TNFRSF6B, PDYN, GH1, OPALIN, SYN1, CAMK2A, NEFH, NEUR0D6 or 0LIG2 promoter.

12. The AAV vector of any one of the preceding claims, wherein the promoter is a CMV, CBh, CBA, CAG, CMVe-GFAP, GFAP, CMVe-0LIG2, 0LIG2 or SYN1 promoter.

13. The AAV vector of any one of the preceding claims, wherein the promoter is a CBh, CMVe-GFAP, CMVe-0LIG2 or a CBA promoter, preferably a CBh or CMVe-GFAP promoter.

14. The AAV vector of any one of the preceding claims, wherein the promoter is selected from a promoter comprising the nucleotide sequence of SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32 or SEQ ID NO: 42 or a promoter having at least 99% identity therewith.

15. The AAV vector of any one of the preceding claims, wherein the IRES comprises the nucleic acid sequence of SEQ ID NO: 33 or 34 or an IRES having at least 99% identity therewith.

16. The AAV vector of any one of the preceding claims, wherein the secretion peptide is a mouse IgG kappa chain secretion peptide, a mouse heavy chain secretion peptide, a human growth hormone 1 secretion peptide, a human heavy chain secretion peptide, a human light chain secretion peptide, a human Oncostatin M (OSM) secretion peptide, a Vesicular stomatitis virus G protein (VSV-G) secretion peptide, a basement-membrane protein 40 (BM40) secretion peptide, a Secrecon secretion peptide, a CD33 secretion peptide, a tissue plasminogen activator (tPA) secretion peptide, a Human Chymotrypsinogen secretion peptide, a Human trypsinogen-2 secretion peptide, a Human IL-2 secretion peptide, a Gaussia luciferase secretion peptide, an Human serum albumin secretion peptide, an Influenza Haemagglutinin secretion peptide, a Human insulin secretion peptide or a Silkworm Fibroin LC secretion peptide.

17. The AAV vector of any one of the preceding claims, wherein the secretion peptide is a mouse IgG kappa chain secretion peptide, a mouse heavy chain secretion peptide, a human growth hormone 1 secretion peptide, a human heavy chain secretion peptide, or a human light chain secretion peptide.

18. The AAV vector of any one of the preceding claims, wherein the secretion peptide is a mouse IgG kappa chain secretion peptide or a human growth hormone 1 secretion peptide.

19. The AAV vector of any one of the preceding claims, wherein the secretion peptide comprises the amino acid sequence of SEQ ID NO: 40 or 41, or a secretion peptide having at least 99% identity therewith.

20. The AAV vector of any one of the preceding claims, wherein the regulatory element comprises a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) and / or at least one miR binding site.

21. The AAV vector of any one of the preceding claims, wherein the regulatory element comprises a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), optionally wherein the WPRE comprises the nucleic acid sequence of SEQ ID NO: 35 or a WPRE having at least 99% identity therewith.

22. The AAV vector of any one of the preceding claims, wherein the regulatory element comprises at least one miR binding site.

23. The AAV vector of any one of the preceding claims, wherein the regulatory element comprises at least one miR- 122 binding site, miR-1 binding site, miR- 142 binding site or miR- 183 binding site.

24. The AAV vector of any one of the preceding claims, wherein the regulatory element comprises at least one, preferably three, miR- 122 binding sites.

25. The AAV vector of any one of claims 20 to 24, wherein the miR binding site comprises the nucleic acid sequence of SEQ ID NO: 36 or a miR binding site having at least 80%, 85%, 90% or 95% identity therewith.

26. The AAV vector of any one of the preceding claims, wherein the expression cassette further comprises a poly-adenylation (poly A) element.

27. The AAV vector of any one of the preceding claims, wherein the AAV vector is a wildtype or engineered AAV vector.

28. The AAV vector of any one of the preceding claims, wherein the AAV vector is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh.8, an AAVrh.9, AAVrh.10, an AAVrh. l l, an AAVrh.12, an engineered AAVrh.13, an AAVrh.14, an AAVrh.15, an AAVrh.16, an AAVrh.17 or an AAV.Hu68.

29. The AAV vector of any one of the preceding claims, wherein the AAV vector is an engineered AAV1, engineered AAV2, an engineered AAV3, an engineered AAV4, an engineered AAV5, an engineered AAV6, an engineered AAV7, an engineered AAV8, an engineered AAV9, an engineered AAV10, an engineered AAV11, an engineered AAV12, an engineered AAVrh.8, an engineered AAVrh.9, engineered AAVrh.10, an engineered AAVrh.11, an engineered AAVrh.12, an engineered AAVrh.13, anengineered AAVrh.14, an engineered AAVrh.15, an engineered AAVrh.16, an engineered AAVrh.17 or an engineered AAV.Hu68.

30. The AAV vector of any one of the preceding claims, wherein the AAV vector is an AAV2-BR1, AAV-S, AAV-F, AAV.PHP.eB, AAV9.PHP.V1, AAV1RX, AAV1R6, AAV1R7, AAV.CAP-B10, AAV.CAP-B22, AAV.CAP-Mac, AAV VCAP-103, AAV bCap 1, V-CAP102, AAV-CGN1, AAV-STRV5, AAV-PAL2, AAV-MDV1A, AAV- MDV1B, AAV-MaCPNSl, AAV-MaCPNS2 or AAV VCAP-100.

31. The AAV vector of any one of the preceding claims, wherein the AAV vector is an AAV9, AAVrh.10, AAV.PHP.eB or AAV2-BR1 vector, preferably an AAV9, AAVrh.10 or AAV.PHP.eB vector.

32. The AAV vector of any one of the preceding claims, wherein the anti-TDP-43 antibody(i) binds to an epitope within amino acids 397-411 of human TDP-43 (SEQ ID NO: 1);(ii) binds physiologically functional and pathological / misfolded TDP-43; (iii) inhibits TDP-43 aggregation; and / or (iv) reduces the level of phosphorylated TDP-43.

33. The AAV vector of any one of the preceding claims, wherein the antibody is a Fab, IgGl or IgGl.

34. The AAV vector of any one of the preceding claims, wherein the anti-TDP-43 antibody is an IgGl or IgG4.

35. The AAV vector of any one of the preceding claims, wherein the anti-TDP-43 antibody comprises an Fc mutation, preferably an S228P mutation.

36. An AAV vector, preferably an AAV9 vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody , (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody , and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising theamino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

37. An AAV vector, preferably an AAV9 vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CMVe-GFAP promoter, (ii) the light chain of an antibody , (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody , and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

38. An AAV vector, preferably an AAV9 vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CMVe-0LIG2 promoter, (ii) the light chain of an antibody , (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody , and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising theamino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

39. An AAV vector, preferably an AAVrh.10 vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody , (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

40. An AAV vector, preferably an AAV.PHP.eB vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising theamino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

41. An AAV vector, preferably an AAV.PHP.eB vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBh promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) three miR-122 binding sites, wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

42. An AAV vector, preferably an AAV.PHP.eB vector, comprising an expression cassette comprising a nucleic acid sequence encoding from 5’ to 3’ : (i) a CBA promoter, (ii) the light chain of an antibody, (iii) an internal ribosome entry site (IRES), (iv) the heavy chain of an antibody, and (v) a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE), wherein the light chain of the antibody comprises a secretion peptide and the heavy chain of the antibody comprises a secretion peptide, and wherein the antibody is an anti-TDP-43 antibody comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequenceof SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

43. The AAV vector of any one of claims 36 to 42, wherein the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO: 12, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

44. The AAV vector of claim 43, wherein the anti-TDP-43 antibody is a murine or chimeric antibody.

45. The AAV vector of claim 43 or 44, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 or a VH having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 14 or a VL having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14.

46. The AAV vector of any one of claims 43 to 45, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 10 and a VL comprising the amino acid sequence of SEQ ID NO: 14.

47. The AAV vector of any one of claims 36 to 42, wherein the anti-TDP-43 antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VH- CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser), a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27.

48. The AAV vector of claim 47, wherein the anti-TDP-43 antibody is a humanized antibody.

49. The AAV vector of claim 47 or 48, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 or a VH having at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a VL comprising the amino acid sequence of SEQ ID NO: 24 or a VL having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24.

50. The AAV vector of any one of claims 47 to 49, wherein the anti-TDP-43 antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 20 and a VL comprising the amino acid sequence of SEQ ID NO: 24.

51. A pharmaceutical composition comprising the AAV vector of any one of the preceding claims and a pharmaceutically acceptable carrier and / or excipient and / or diluent.

52. The AAV vector of any one of claims 1 to 50, or the pharmaceutical composition of claim 51, for use in a method of reducing the level of phosphorylated TDP-43 in a subject.

53. The AAV vector or pharmaceutical composition for use of claim 52, wherein the level of phosphorylated TDP-43 is reduced in the hippocampus of the subject.

54. The AAV vector of any one of claims 1 to 50, or the pharmaceutical composition of claim 51, for use in prevention, alleviation or treatment of a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy.

55. The AAV vector or the pharmaceutical composition for use according to claim 54, wherein the disease disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy is Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD).

56. The AAV vector or the pharmaceutical composition for use according to claim 54 or 55, wherein the disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), or limbic-predominant age-related TDP-43 encephalopathy (LATE).

57. The AAV vector or the pharmaceutical composition for use according to any one of claims 54 to 56, wherein the disease, disorder and / or abnormality associated with TDP- 43, or TDP-43 proteinopathy, is Alzheimer’s disease (AD).

58. The AAV vector or the pharmaceutical composition for use according to any one of claims 54 to 56, wherein the disease, disorder and / or abnormality associated with TDP- 43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD).

59. The AAV vector or the pharmaceutical composition for use according to any one of claims 54 to 56, wherein the disease, disorder and / or abnormality associated with TDP- 43, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

60. A method of retaining or increasing cognitive memory capacity or slowing memory loss in a subject with a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, comprising administering the AAV vector of any one of claims 1 to 50 or the pharmaceutical composition of claim 51 to the subject.

61. A method of reducing the level of aggregated TDP-43 and / or phosphorylated TDP-43 in a subject, comprising administering the AAV vector of any one of claims 1 to 50 or pharmaceutical composition of claim 51 to the subject.

62. The method of claim 61, wherein the level of aggregated TDP-43 and / or phosphorylated TDP-43 is reduced in the hippocampus of the subject.

63. The method of claim 61 or 62, wherein the method comprises administering at least one additional therapeutic agent.

64. The method of claim 63, wherein the additional therapeutic agent targets alpha- synuclein, BACE1, Tau, beta-amyloid, TDP-43 or a neuroinflammation protein.

65. A method of inducing production of an anti-TDP-43 antibody in a subject, comprising administering the AAV vector of any one of claims 1 to 50 or the pharmaceutical composition of claim 51 to the subject.

66. A method of inducing sustained anti-TDP-43 antibody expression in a subject, comprising administering the AAV vector of any one of claims 1 to 50 or pharmaceutical composition of claim 51 to the subject.

67. The method of claim 65 or 66, wherein the concentration of the anti-TDP-43 antibody in the blood plasma or serum of the subject is from 1 pg / mL to 1000 pg / mL after the administration.

68. The method of any one of claims 66 to 67, wherein the blood plasma or serum level of the anti-TDP-43 antibody is sustained at a concentration of at least 50 pg / mL, at least 100 pg / mL, or at least 200 pg / mL for a period of at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 16 weeks after the administration.

69. The method of any one of claims 65 to 68, wherein the concentration of the anti-TDP-43 antibody in the CSF of the subject is from 1 ng / mL to 1000 ng / mL after the administration.

70. The method of any one of claims 66 to 69, wherein the CSF level of the anti-TDP-43 antibody is sustained at a concentration of at least 50 ng / mL, at least 100 ng / mL, or atleast 200 ng / mL for a period of at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 16 weeks after the administration.

71. The method of any one of claims 66 to 70, wherein the sustained concentration of anti- TDP-43 antibody is achieved after a single administration.

72. The AAV vector or pharmaceutical composition for use of any one of claims 52 to 59 or the method of any one of claims 60 to 71, wherein the anti-TDP-43 antibody is expressed in the parenchyma, preferably in multiple brain regions.

73. The AAV vector or pharmaceutical composition for use or the method of claim 72, wherein the multiple brain regions are selected from two or more of the isocortex, hippocampus and cerebellum.

74. The AAV vector or pharmaceutical composition for use of any one of claims 52 to 59, 72 or 73, or the method of any one of claims 60 to 73, wherein the anti-TDP-43 antibody is expressed with a mean CSF to serum ratio or CSF to plasma ratio of greater than 0.1%, 0.2% or more, 0.5% or more, 1% or more, 2% or more, 3% or more, 4% or more, 4.5% or more, or 5% or more.

75. The AAV vector or pharmaceutical composition for use according to any one of claims 52 to 59 or 72 to 74, or the method according to any one of claims 60 to 74, wherein the AAV vector is administered by parenteral route.

76. The AAV vector or pharmaceutical composition for use, or the method according to claim 75, wherein the AAV vector is administered by intravenous, intraperitoneal, intranasal, intravitreous, subcutaneous, intramuscular, intrathecal, intracisternal, intraparenchymal, intrastriatal or intracerebroventricular route.

77. The AAV vector or pharmaceutical composition for use according to any one of claims 52 to 59, or 72 to 76, or the method according to any one of claims 60 to 76, wherein the AAV vector administration comprises delivery of from 1 • 1010vg / kg to 5 • 1014vg / kg of the AAV vector to the subject per dose.

78. A method for producing an AAV vector, comprising: (i) providing a host cell comprising the viral genome of the AAV vector of any one of claims 1 to 50; (ii) incubating the host cell under conditions suitable to enclose the viral genome in an AAV capsid, thereby making the AAV vector.